A synthetic cDNA for human CYP17A1 was modified to delete residues 2–19, substitute the hydrophilic sequence 20RRCP23 (link) with 20AKKT23 (link), and add a C-terminal four histidine tag (fig. S6 ) before cloning into the pCWori+ plasmid and overexpression in E. coli JM109 cells. Protein was purified by nickel affinity, cation exchange, and size exclusion chromatography. Abiraterone was synthesized (Methods). Binding affinities were determined using a UV/vis spectral shift assay. Progesterone 17α-hydroxylation was evaluated using HPLC separation and UV detection. For crystallography, inhibitors were included throughout purification. Crystals were grown from CYP17A1 (30 mg/mL) complexed with inhibitor using hanging-drop vapor diffusion to equilibrate against 30% PEG 3350, 0.175 M Tris, pH 8.5, 0.30 M ammonium sulfate, and 3% glycerol. Diffraction data was collected and phased by molecular replacement. Iterative model building and refinement generated the final model. Substrates were docked using Surflex-Dock30 (link).
>
Chemicals & Drugs
>
Inorganic Chemical
>
Ammonium nickel sulfate
Ammonium nickel sulfate
Ammonium nickel sulfate is a chemical compound with the formula (NH4)2Ni(SO4)2.
It is a green crystalline solid that is soluble in water and used in various industrial and research applications.
As a metal salt, ammonium nickel sulfate has applications in electroplating, pigments, and as a precursor for other nickel compounds.
In scientific research, it may be employed as a source of nickel ions for studies involving metal-based catalysts, biochemical reactions, or materials science.
The chemical and physical properties of ammonium nickel sulfate make it a versatile tool for researchers across multple disciplines.
It is a green crystalline solid that is soluble in water and used in various industrial and research applications.
As a metal salt, ammonium nickel sulfate has applications in electroplating, pigments, and as a precursor for other nickel compounds.
In scientific research, it may be employed as a source of nickel ions for studies involving metal-based catalysts, biochemical reactions, or materials science.
The chemical and physical properties of ammonium nickel sulfate make it a versatile tool for researchers across multple disciplines.
Most cited protocols related to «Ammonium nickel sulfate»
abiraterone
Biological Assay
Cells
Crystallography
CYP17A1 protein, human
Diffusion
DNA, Complementary
Escherichia coli
Gel Chromatography
Glycerin
High-Performance Liquid Chromatographies
Histidine
Homo sapiens
Hydroxylation
inhibitors
Nickel
Plasmids
polyethylene glycol 3350
Progesterone
Proteins
Sulfate, Ammonium
Tromethamine
Protein Expression, Purification, and Crystallization—The gene encoding single chain (VH-linker-VL) antibody 80R (scFv) was cloned into pET22b (Novagen) containing an N-terminal periplasmic secretion signal pelB, and a thrombin-removable C-terminal His6 tag. 80R was overexpressed in BL21(DE3) cells at 30 °C for 15 h with 1 m
The gene encoding S1-RBD (residues 318-510) was cloned into vector pAcGP67A (Pharmingen) containing an N-terminal gp67 secretion signal and a thrombin-cleavable C-terminal His6 tag. It was expressed in Sf9 cells (Invitrogen) with a multiplicity of infection = 5 for 72 h. Similar to 80R, S1-RBD was purified from the media with HisBind nickel-nitrilotriacetic acid and Superdex 200 columns, with thrombin digestion. N-Linked glycosylation was removed by incubation with peptide:N-glycosidase F (New England Biolabs) at 23 °C, as monitored by SDS-PAGE. S1 RBD-80R complexes were formed by mixing the two purified components and isolated by gel filtration with Superdex 200 in 10 m
Crystals grew by the hanging drop vapor diffusion method at 17 °C over ∼21 days. For S1-RBD, 2 μl of S1-RBD was mixed with an equal volume of well solution containing 4% w/v polyethylene glycol 4000, 0.1
Data Collection, Structure Determination, and Refinement— X-ray diffraction data were collected at the National Synchrotron Light Source beamline X6A and X29A for S1-RBD crystals, the Stanford Synchrotron Radiation Laboratory beamline 11.1, and at the Advanced Light Source beamlines 5.0.3 and 12.3.1 for crystals of the S1-RBD-80R complex. Glycerol (25%) was used as a cryoprotectant in both cases. All the data were processed with DENZO and SCALEPACK or with the HKL2000 package (8 ). Crystals of S1 RBD adopt space group P43212 with unit cell dimensions a = 75.9 and c = 235.8 (
Data collection and refinement statistics
| Cell parameters | a = 75.9, c = 235.9 Å | a = 47.5, b = 175.9, c = 67.6 Å; β = 96.6° |
| Space group | P43212 | P21 |
| Resolution (Å) | 2.2 | 2.3 |
| Total reflections | 233011 | 159047 |
| Unique reflections | 36036 | 51915 |
| Completeness (%) | 99.9 (99.9) | 93.8 (87.0) |
| Average I/σ(I) | 24.7 (2.0) | 8.8 (1.9) |
| Rmerge | 0.098 (0.739) | 0.145 (0.571) |
| Redundancy | 6.5 | 3.1 |
| Rwork | 0.182 (0.230) | 0.248 (0.301) |
| Rfree (5% data) | 0.213 (0.289) | 0.295 (0.391) |
| r.m.s.d. bond distance (Å) | 0.013 | 0.009 |
| r.m.s.d. bond angle (°) | 1.49 | 1.22 |
| Average B value | 50.0 | 37.1 |
| Solvent atoms | 152 | 470 |
| Residues in most favored regions | 276 | 631 |
| Residues in additional allowed regions | 35 | 81 |
| Residues in generously allowed regions | 3 | 5 |
| Residues in disallowed regions | 0 | 0 |
Numbers in parentheses correspond to the highest resolution shell (2.28-2.20 Å for S1 RBD; 2.29-2.38 Å for S1 RBD-80R)
Numbers in parentheses correspond to the highest resolution shell (2.26-2.20 Å for S1 RBD; 2.29-2.38 Å for S1 RBD-80R)
r.m.s.d., root meant square deviation
where A and B were obtained by least square fitting of the averaged measured intensities. The ratio of the parameters B and A (B/A = 0.65) coincided with the height ratio of the Patterson peak at (1/3, 0, 0), as required by the lattice-translocation theory presented by Wang. The corrected intensity distribution (
The structure of uncomplexed S1-RBD (which showed no lattice defects) was determined by molecular replacement with PHASER (13 (link)) using S1-RBD from the structure of the S1-RBD-ACE2 complex (Protein Data Bank code 2AJF) as the search model. The asymmetric unit contains two molecules of S1-RBD arranged as a symmetric dimer. The final model includes residues 320-503 of both monomers and 152 water molecules.
Geometric parameters are excellent as assessed with PRO-CHECK (18 ) (
The cytoarchitecture of the PFC was studied in ten adult, male mice (strain C57BL/6) of similar weight (approximately 20 g). These control mouse brains were kindly donated and immersion fixed by Dr. H. Manji, NIMH, USA. All animal procedures were in strict accordance with the NIH animal care guidelines. The histological processing of these brains was performed at the laboratory of Dr. Rajkowska. The brains were embedded in 12% celloidin, cut into 40-μm serial sections using a sliding microtome and Nissl (1% cresyl violet) stained. Celloidin was chosen as an embedding medium to allow for the preparation of ‘thick’ sections with clear morphology and high contrast of Nissl-stained neurons and glial cells. In these immersion-fixed brains, any spots showing pycnotic reaction were not incorporated in this study.
In addition to these ten mice, four adult male mice (C57BL/6 strain) were stained for dopamine and four adult male mice for AChE, myelin, and immunohistochemically for SMI, PV and CB. For each staining, a different set of sections with several consecutive sections stained with Nissl at HBMU’s laboratory was used. The antibodies applied were the dopamine (DA) antibody (Geffard et al. 1984 (link)), SMI-32 antibody (Sternberger Monoclonals Inc., Baltimore, MD, USA: monoclonal antibody to one epitope of non-phosphorylated tau neurofilaments, lot number 11), SMI-311antibody (pan-neuronal neurofilament marker cocktail of several monoclonal antibodies for several epitopes of non-phosphorylated tau protein, Sternberger Monoclonals Inc., Baltimore, MD, USA: lot number 9) (SMI antibodies are presently distributed through Covance Research Products, USA), monoclonal anti-CB D-28K antibody (Sigma, St. Louis, MO, USA: product number C-9848, clone number CB-955, lot number 015K4826), and monoclonal anti-PV antibody (Sigma, St. Louis, MO, USA: product number P-3171, clone number PA-235, lot number 026H4824). Mice to be stained for DA were intracardially perfused under deep pentobarbital anesthesia (1 ml/kg body weight, i.p.), with saline followed by fixative. For DA staining, the fixative was 5% glutaraldehyde in 0.05 M acetate buffer at pH 4.0. After perfusion, the brains were immersed in 0.05 Tris containing 1% sodium disulfite (Na2S2O5) at pH 7.2 (De Brabander et al. 1992 (link)). Mouse PFC was sectioned at 40 μm by a vibratome. These sections were stained overnight in a cold room at 4°C using the polyclonal primary antibody sensitive to DA that was raised in the Netherlands Institute for Brain Research (NIBR) (Geffard et al. 1984 (link)), the specificity of which had been demonstrated previously (Kalsbeek et al. 1990 (link)). DA antiserum was diluted 1:2,000 in 0.05 M Tris containing 1% Na2S2O5 and 0.5% Triton X-100, pH 7.2. After overnight incubation, the sections were washed three times with Tris-buffered saline (TBS) and subsequently incubated in the secondary antibody goat–antirabbit, also raised in NIBR at 1:100 for 1 h. After having been rinsed 3× in TBS, it was incubated in the tertiary antibody, peroxidase–antiperoxidase, at 1:1,000 for 60 min. Both the secondary and the tertiary antibodies were diluted in TBS with 0.5% gelatine and 0.5% Triton X-100. For visualization, the sections were transferred into 0.05% diaminobenzidine (DAB; Sigma) with 0.5% nickel ammonium sulfate. The reaction was stopped after a few minutes by transferring the sections to TBS (3 × 10 min), then the sections were mounted on slides, air dried, washed, dehydrated and coverslipped.
Mice to be stained with anti-PV, anti-CB and SMI-32 and SMI-311 were fixed with 4% formaldehyde solution in 0.1 M phosphate buffer at pH 7.6. Mouse PFC was sectioned at 40 μm by a vibratome. To prevent endogenous peroxidase activity, free-floating sections were pretreated for 30 min in a Tris-buffered saline (TBS) solution containing 3% hydrogen peroxide and 0.2% Triton X-100. To prevent non-specific antibody staining, these sections were placed in a milk solution (TBS containing 5% nonfat dry milk and 0.2% Triton X-100) for 1 h. Incubation of the primary antibody, directly after the milk step was carried out overnight in a cold room at 4°C. The primary antibodies were diluted in the above-mentioned milk solution: SMI-32 and SMI-311 at 1:1,000, PV antibody at 1:1,000, and CB antibody at 1:250. For the monoclonal SMI-32, SMI-311, PV and CB antibodies, raised in mice, we used peroxidase-conjugated rabbit–antimouse (1:100 in 5% milk solution with 0.2% Triton X-100) as a secondary antibody. Visualization took place in 0.05% diaminobenzidine enhanced with 0.2% nickel ammonium sulfate. The reaction was stopped after a few minutes by transferring these sections to TBS (3 × 10 min), after which the sections were rinsed in distilled water, mounted on slides, air dried, washed, dehydrated and coverslipped. Control sections that were incubated according to the same procedure as described above, omitting the primary antibody, were all negative. All sections were cut coronally, because the coronal plane offers in general the best view to differentiate between the subareas of the rodent PFC (Uylings et al. 2003 (link); Van de Werd and Uylings 2008 (link)).
Sections were processed for AChE staining according to the protocol described by Cavada et al. (1995 (link)). The sections were incubated overnight in a solution of cupric sulfate and acetate buffer at pH 5 to which acetylthiocholine iodide and ethopropazine were added just before the start of incubation. After rinsing, the sections were developed in a sodium sulfide solution until a light brown color appeared and subsequently intensified to a dark brown color in a silver nitrate solution. Finally, the sections were differentiated after rinsing in a thiosulfate solution, dehydrated and mounted. In all steps, the solutions and sections were shaken constantly. The myelin was stained with silver by physical development according to Gallyas (1979 (link)). The sections were first placed in 100% ethanol and then immersed in a 2:1 solution of pyridine and acetic acid for 30 min. After rinsing, they were placed in an ammonium silver nitrate solution and after rinsing with 0.5% acetic acid, the sections were immersed in the optimal physical developer solution at room temperature (Gallyas 1979 (link)) until they showed good stain intensity under the microscope. Then the development of the staining was stopped in 0.5% acetic acid and the sections were dehydrated and mounted with Histomount. The sections were studied at intervals of 80–160 μm, and examined under a light microscope at a 63× magnification.
In addition to these ten mice, four adult male mice (C57BL/6 strain) were stained for dopamine and four adult male mice for AChE, myelin, and immunohistochemically for SMI, PV and CB. For each staining, a different set of sections with several consecutive sections stained with Nissl at HBMU’s laboratory was used. The antibodies applied were the dopamine (DA) antibody (Geffard et al. 1984 (link)), SMI-32 antibody (Sternberger Monoclonals Inc., Baltimore, MD, USA: monoclonal antibody to one epitope of non-phosphorylated tau neurofilaments, lot number 11), SMI-311antibody (pan-neuronal neurofilament marker cocktail of several monoclonal antibodies for several epitopes of non-phosphorylated tau protein, Sternberger Monoclonals Inc., Baltimore, MD, USA: lot number 9) (SMI antibodies are presently distributed through Covance Research Products, USA), monoclonal anti-CB D-28K antibody (Sigma, St. Louis, MO, USA: product number C-9848, clone number CB-955, lot number 015K4826), and monoclonal anti-PV antibody (Sigma, St. Louis, MO, USA: product number P-3171, clone number PA-235, lot number 026H4824). Mice to be stained for DA were intracardially perfused under deep pentobarbital anesthesia (1 ml/kg body weight, i.p.), with saline followed by fixative. For DA staining, the fixative was 5% glutaraldehyde in 0.05 M acetate buffer at pH 4.0. After perfusion, the brains were immersed in 0.05 Tris containing 1% sodium disulfite (Na2S2O5) at pH 7.2 (De Brabander et al. 1992 (link)). Mouse PFC was sectioned at 40 μm by a vibratome. These sections were stained overnight in a cold room at 4°C using the polyclonal primary antibody sensitive to DA that was raised in the Netherlands Institute for Brain Research (NIBR) (Geffard et al. 1984 (link)), the specificity of which had been demonstrated previously (Kalsbeek et al. 1990 (link)). DA antiserum was diluted 1:2,000 in 0.05 M Tris containing 1% Na2S2O5 and 0.5% Triton X-100, pH 7.2. After overnight incubation, the sections were washed three times with Tris-buffered saline (TBS) and subsequently incubated in the secondary antibody goat–antirabbit, also raised in NIBR at 1:100 for 1 h. After having been rinsed 3× in TBS, it was incubated in the tertiary antibody, peroxidase–antiperoxidase, at 1:1,000 for 60 min. Both the secondary and the tertiary antibodies were diluted in TBS with 0.5% gelatine and 0.5% Triton X-100. For visualization, the sections were transferred into 0.05% diaminobenzidine (DAB; Sigma) with 0.5% nickel ammonium sulfate. The reaction was stopped after a few minutes by transferring the sections to TBS (3 × 10 min), then the sections were mounted on slides, air dried, washed, dehydrated and coverslipped.
Mice to be stained with anti-PV, anti-CB and SMI-32 and SMI-311 were fixed with 4% formaldehyde solution in 0.1 M phosphate buffer at pH 7.6. Mouse PFC was sectioned at 40 μm by a vibratome. To prevent endogenous peroxidase activity, free-floating sections were pretreated for 30 min in a Tris-buffered saline (TBS) solution containing 3% hydrogen peroxide and 0.2% Triton X-100. To prevent non-specific antibody staining, these sections were placed in a milk solution (TBS containing 5% nonfat dry milk and 0.2% Triton X-100) for 1 h. Incubation of the primary antibody, directly after the milk step was carried out overnight in a cold room at 4°C. The primary antibodies were diluted in the above-mentioned milk solution: SMI-32 and SMI-311 at 1:1,000, PV antibody at 1:1,000, and CB antibody at 1:250. For the monoclonal SMI-32, SMI-311, PV and CB antibodies, raised in mice, we used peroxidase-conjugated rabbit–antimouse (1:100 in 5% milk solution with 0.2% Triton X-100) as a secondary antibody. Visualization took place in 0.05% diaminobenzidine enhanced with 0.2% nickel ammonium sulfate. The reaction was stopped after a few minutes by transferring these sections to TBS (3 × 10 min), after which the sections were rinsed in distilled water, mounted on slides, air dried, washed, dehydrated and coverslipped. Control sections that were incubated according to the same procedure as described above, omitting the primary antibody, were all negative. All sections were cut coronally, because the coronal plane offers in general the best view to differentiate between the subareas of the rodent PFC (Uylings et al. 2003 (link); Van de Werd and Uylings 2008 (link)).
Sections were processed for AChE staining according to the protocol described by Cavada et al. (1995 (link)). The sections were incubated overnight in a solution of cupric sulfate and acetate buffer at pH 5 to which acetylthiocholine iodide and ethopropazine were added just before the start of incubation. After rinsing, the sections were developed in a sodium sulfide solution until a light brown color appeared and subsequently intensified to a dark brown color in a silver nitrate solution. Finally, the sections were differentiated after rinsing in a thiosulfate solution, dehydrated and mounted. In all steps, the solutions and sections were shaken constantly. The myelin was stained with silver by physical development according to Gallyas (1979 (link)). The sections were first placed in 100% ethanol and then immersed in a 2:1 solution of pyridine and acetic acid for 30 min. After rinsing, they were placed in an ammonium silver nitrate solution and after rinsing with 0.5% acetic acid, the sections were immersed in the optimal physical developer solution at room temperature (Gallyas 1979 (link)) until they showed good stain intensity under the microscope. Then the development of the staining was stopped in 0.5% acetic acid and the sections were dehydrated and mounted with Histomount. The sections were studied at intervals of 80–160 μm, and examined under a light microscope at a 63× magnification.
We estimated population-level exposures for different groups (e.g., race/ethnicity) to PM2.5 and for the following 14 PM2.5 components measured by the U.S. EPA’s national monitoring network: sulfate (SO42–), nitrate (NO3–), ammonium (NH4+), organic carbon matter (OCM), elemental carbon (EC), sodium ion (Na+), aluminum (Al), calcium (Ca), chlorine (Cl), nickel (Ni), silicon (Si), titanium (Ti), vanadium (V), and zinc (Zn). These components were selected because they contribute ≥ 1% to total PM2.5 mass for yearly or seasonal averages, and/or have been associated with adverse health outcomes in previous studies including mortality, heart rate, heart rate variability, and low birth weight (Bell et al. 2007 (link), 2009 (link); Dominici et al. 2007 (link); Franklin et al. 2008 (link); Huang et al. 2012 (link); Lippmann et al. 2006 (link); Ostro et al. 2007 (link), 2008 (link); Rohr et al. 2011 (link); Wilhelm et al. 2012 (link)).
Daily air pollution measures were obtained for 2000 through 2006 (U.S. EPA 2011a ). Pollutant monitors were matched to U.S. census tracts, which are geographic units representing small subdivisions of a county and are the smallest spatial unit for which demographic variables of interest were available. Tracts from the 2000 Census (U.S. Census Bureau 2007 ) were designed to have an optimal population of 4,000 persons (range, 1,500–8,000) and to follow government boundaries (e.g., county), geographic features (e.g., rivers), or other identifiable features (e.g., roadways), where possible. The median land area of the 2000 census tracts in the continental United States was 5.06 km2.
Census tracts in the continental United States were included in our analysis if they had PM2.5 component monitors in operation for ≥ 3 years with ≥ 180 days of observations during the study period. Results were based on 219 monitors in 215 census tracts. Land use near monitors was 43% residential, 34% commercial, 8% industrial, 8% agricultural, and 4% forest.
We calculated long-term averages for each pollutant and 2000 census tract with a monitor for that pollutant. If multiple monitors were present for the same pollutant in a single tract, we averaged daily monitor values within a tract, and then averaged daily values to generate long-term averages. The population and area of census tracts varied. The mean (± SD) distance between a census tract’s centroid and monitor was 2.3 km ± 4.9 km (median 0.8 km; maximum 46.7 km).
For each census tract, we considered population characteristics (U.S. Census 2007 ):
We excluded census tracts with populations ≤ 100 (n = 1; for tract with population = 1). For each population characteristic and category (e.g., race/ethnicity, Hispanic), we estimated the average exposure to each pollutant for that group in the United States as a whole by weighting levels in each census tract by the population as
where Yik is the national average estimated exposure to pollutant k for persons with characteristic i (e.g., Hispanic), j is the number of census tracts with pollutant data (J = 215), Pi,j is the number of persons with characteristic i in census tract j, and xjk is the concentration of pollutant k for census tract j. This provides an estimate of average exposure for each pollutant and population group, accounting for population size and pollutant levels in each census tract. In addition, we performed univariate regression to estimate differences in exposure to PM2.5 and for each component according to census tract characteristics (e.g., percentage of persons unemployed), which are expressed as the percent change in exposure compared with overall mean levels associated with a 10% increase in a given population characteristic.
Whereas the regression analysis investigated whether some groups had higher exposures than others among areas with monitors, we further contrasted population characteristics between census tracts with and without monitors for PM2.5 or its components. We calculated population characteristics for census tracts with and without monitors and performed univariate logistic regression to estimate the percent increase in the probability of a census tract having a monitor with a 10% increase in each population characteristic. This analysis investigated whether some populations are better covered by the existing monitoring network than others.
Daily air pollution measures were obtained for 2000 through 2006 (U.S. EPA 2011a ). Pollutant monitors were matched to U.S. census tracts, which are geographic units representing small subdivisions of a county and are the smallest spatial unit for which demographic variables of interest were available. Tracts from the 2000 Census (U.S. Census Bureau 2007 ) were designed to have an optimal population of 4,000 persons (range, 1,500–8,000) and to follow government boundaries (e.g., county), geographic features (e.g., rivers), or other identifiable features (e.g., roadways), where possible. The median land area of the 2000 census tracts in the continental United States was 5.06 km2.
Census tracts in the continental United States were included in our analysis if they had PM2.5 component monitors in operation for ≥ 3 years with ≥ 180 days of observations during the study period. Results were based on 219 monitors in 215 census tracts. Land use near monitors was 43% residential, 34% commercial, 8% industrial, 8% agricultural, and 4% forest.
We calculated long-term averages for each pollutant and 2000 census tract with a monitor for that pollutant. If multiple monitors were present for the same pollutant in a single tract, we averaged daily monitor values within a tract, and then averaged daily values to generate long-term averages. The population and area of census tracts varied. The mean (± SD) distance between a census tract’s centroid and monitor was 2.3 km ± 4.9 km (median 0.8 km; maximum 46.7 km).
For each census tract, we considered population characteristics (U.S. Census 2007 ):
We excluded census tracts with populations ≤ 100 (n = 1; for tract with population = 1). For each population characteristic and category (e.g., race/ethnicity, Hispanic), we estimated the average exposure to each pollutant for that group in the United States as a whole by weighting levels in each census tract by the population as
where Yik is the national average estimated exposure to pollutant k for persons with characteristic i (e.g., Hispanic), j is the number of census tracts with pollutant data (J = 215), Pi,j is the number of persons with characteristic i in census tract j, and xjk is the concentration of pollutant k for census tract j. This provides an estimate of average exposure for each pollutant and population group, accounting for population size and pollutant levels in each census tract. In addition, we performed univariate regression to estimate differences in exposure to PM2.5 and for each component according to census tract characteristics (e.g., percentage of persons unemployed), which are expressed as the percent change in exposure compared with overall mean levels associated with a 10% increase in a given population characteristic.
Whereas the regression analysis investigated whether some groups had higher exposures than others among areas with monitors, we further contrasted population characteristics between census tracts with and without monitors for PM2.5 or its components. We calculated population characteristics for census tracts with and without monitors and performed univariate logistic regression to estimate the percent increase in the probability of a census tract having a monitor with a 10% increase in each population characteristic. This analysis investigated whether some populations are better covered by the existing monitoring network than others.
Full text: Click here
Alexa 350
Alexa594
alexa fluor 488
ammonium nickel sulfate
Antibodies
Antibodies, Anti-Idiotypic
Avidin
azo rubin S
Biopharmaceuticals
Biotin
Cloning Vectors
Diploid Cell
Equus asinus
Goat
HCRT protein, human
Hypothalamus
Immunohistochemistry
Mice, House
Microscopy
POU3F2 protein, human
Rabbits
Streptavidin
Vascular Access Ports
Most recents protocols related to «Ammonium nickel sulfate»
All of the reagents and materials were noted to be of analytical quality. Vancomycin (≥85%, CAS no. 1404-93-9), tetra-chloroauric acid trihydrate (HAuCl4.3H2O; 99.9%, CAS no. 16961-25-4), polyethyleneimine (50% w/v in H2O; Mw. 750k, CAS no. 9002-98-6), glutathione, BSA (Bovine serum albumin) and formaldehyde (36.0% in H2O) were obtained from Sigma Aldrich (Mumbai, India). Nickel sulfate, mercuric chloride (99.5%), potassium chloride (99%), Nickel sulfate, magnesium chloride, sodium chloride (99%), ammonium chloride, tryptophan, tyrosine, and solvents were purchased from Merck Life Sciences (Bangalore, India). The other plasticware and glassware were acquired from Tarson (Mumbai, India).
Full text: Click here
Materials and reagents used for the preparation of NiMn-LDH film including Nickel chloride hexahydrate (NiCl2·6H2O, 99%), manganese(ii ) sulfate monohydrate (MnSO4·H2O, 99%), Urea (NH2CONH2, 99%), ammonium fluoride (NH4F, 98%). All chemicals were used without further modifications.
Potassium hydroxide (KOH), potassium persulfate (K2S2O8), nickel sulfate hexahydrate (NiSO4·6H2O), copper nitrate gerhardite (Cu(NO3)2·3H2O), urea (CO(NH2)2), and ammonium hydroxide (NH3·H2O) were purchased from Chongqing Chuandong Co., Ltd. (Chongqing, China). The carbon cloth (type: HCP331N) was obtained from Shanghai Hesen Co., Ltd. (Shanghai, China). All the chemicals were used without further purification.
Full text: Click here
Antibodies used in this study, their manufacturer, specificity, and staining protocol are presented in supplementary tables (Table S2 and S3 ). In brief, after deparaffinization and rehydration, in which sections were washed in xylene (2×10 min) and then in ethanol (100%, 100%, 96%, 90%, 70%, 50%, each×5 min), the sections were washed, treated for antigen retrieval and/or pre-incubated to block background staining. Subsequently, the sections were incubated with primary antibody, then with biotinylated secondary antibody (1:400; Vector Labs), followed by avidin-biotin-peroxidase complex (ABC; 1:800; Vector Labs). Finally, sections were incubated in a solution of 0.5 mg/mL 3, 3-diaminobenzidine (Sigma) in a total volume of 15 mL TBS containing 5 µL H2O2 30% (Merck) and 0.035 g ammonium nickel sulfate (DAB-Ni), at room temperature for color development. For double labeling (DAB-Ni vs. DAB), ammonium nickel sulphate was not included in the second DAB development. The enzyme reaction was stopped in distilled water. Subsequently, the sections were dehydrated, cleared, and coverslipped with Entellan (Merck).
Full text: Click here
Due to the addition of histidine purification tags at both ends of the target gene, the target protein can be affinity adsorbed with nickel chloride or nickel sulfate in the Ni column, and the concentrated fermentation supernatant of A. niger C112-gcdh was separated and purified by using a Ni-NTA column (Sangon Biotech). The concentration and purification process was as follows: 80% saturated ammonium sulfate (dissolved in 20 times) was added to the fermentation supernatant of A. niger C112-gcdh in an ice bath, and the precipitate was taken by centrifugation at 4°C, 10,000 rpm, and 10 min after being stored at 4°C for 12 h. The precipitate was re-dissolved with appropriate amount of pH 7.5 PBS solution and the supernatant was centrifuged at 4°C, 6000 rpm, 5 min. The supernatant was dialyzed in pH 7.5 PBS solution at 4°C for 12 h. After dialysis, the target protein gCDH was purified by Ni-NTA column (which can be stored at-80°C). And target protein was detected by 12% SDS-PAGE (Table 3 ).
Full text: Click here
Top products related to «Ammonium nickel sulfate»
Sourced in United States, Germany, United Kingdom, Italy, China, Japan, Canada, Sao Tome and Principe, Denmark, France, Macao, Australia, Spain, Switzerland
3,3'-diaminobenzidine is a chemical compound commonly used as a chromogenic substrate in various laboratory techniques, such as immunohistochemistry and enzyme-linked immunosorbent assays (ELISA). It is a sensitive and specific reagent that can be used to detect the presence of target proteins or enzymes in biological samples.
Sourced in United States, United Kingdom, Canada, Germany, France, Japan, Switzerland
The Vectastain Elite ABC kit is a specialized laboratory equipment used for the detection and visualization of target proteins or antigens in biological samples. It utilizes an avidin-biotin complex (ABC) system to amplify the signal, enabling researchers to achieve high sensitivity and consistent results in their immunohistochemical or immunocytochemical analyses.
Sourced in United States, Germany, United Kingdom, India, Italy, France, Spain, China, Canada, Sao Tome and Principe, Poland, Belgium, Australia, Switzerland, Macao, Denmark, Ireland, Brazil, Japan, Hungary, Sweden, Netherlands, Czechia, Portugal, Israel, Singapore, Norway, Cameroon, Malaysia, Greece, Austria, Chile, Indonesia
NaCl is a chemical compound commonly known as sodium chloride. It is a white, crystalline solid that is widely used in various industries, including pharmaceutical and laboratory settings. NaCl's core function is to serve as a basic, inorganic salt that can be used for a variety of applications in the lab environment.
Sourced in United States, Canada, United Kingdom, Japan
The ABC Elite kit is a comprehensive laboratory tool designed for versatile applications. It includes a range of high-quality components and accessories to support various experimental procedures. The core function of the kit is to provide researchers with a reliable and efficient solution for their laboratory needs.
Sourced in United States, United Kingdom, Germany, Canada, Italy, Norway
Biotinylated secondary antibody is a laboratory reagent used in various immunoassay techniques. It serves as a detection tool, binding to the primary antibody to amplify the signal and enhance the visualization of target molecules.
Sourced in United States
Nickel ammonium sulfate is a chemical compound with the formula NiSO4(NH4)2SO4·6H2O. It is a green crystalline solid that is used as a laboratory reagent and in various industrial applications.
Sourced in United States, Canada, Japan
The ABC Elite is a versatile laboratory instrument designed for a range of applications. It features advanced technology and precise control mechanisms to ensure reliable and consistent results. The core function of the ABC Elite is to perform essential tasks required in a research or testing environment.
Sourced in United States, United Kingdom, Canada
The Avidin-biotin complex is a high-affinity interaction between the protein avidin and the small molecule biotin. This complex forms the basis for various biotechnological and biochemical applications, enabling the specific and selective detection, separation, or immobilization of biomolecules.
Sourced in Germany, United States, India, United Kingdom, Italy, China, Spain, France, Australia, Canada, Poland, Switzerland, Singapore, Belgium, Sao Tome and Principe, Ireland, Sweden, Brazil, Israel, Mexico, Macao, Chile, Japan, Hungary, Malaysia, Denmark, Portugal, Indonesia, Netherlands, Czechia, Finland, Austria, Romania, Pakistan, Cameroon, Egypt, Greece, Bulgaria, Norway, Colombia, New Zealand, Lithuania
Sodium hydroxide is a chemical compound with the formula NaOH. It is a white, odorless, crystalline solid that is highly soluble in water and is a strong base. It is commonly used in various laboratory applications as a reagent.
Sourced in United States, Germany, United Kingdom, Italy, India, France, Spain, China, Belgium, Sao Tome and Principe, Canada, Denmark, Poland, Australia, Ireland, Israel, Singapore, Macao, Switzerland, Brazil, Mexico, Hungary, Netherlands, Egypt, Japan, Sweden, Indonesia, Czechia, Chile
Potassium chloride (KCl) is an inorganic compound that is commonly used as a laboratory reagent. It is a colorless, crystalline solid with a high melting point. KCl is a popular electrolyte and is used in various laboratory applications.
More about "Ammonium nickel sulfate"
Ammonium nickel sulfate, also known as nickel ammonium sulfate, is a versatile chemical compound with the formula (NH4)2Ni(SO4)2.
This green crystalline solid is soluble in water and has a wide range of industrial and research applications.
As a metal salt, ammonium nickel sulfate is used in electroplating, pigments, and as a precursor for other nickel compounds.
In scientific research, it is often employed as a source of nickel ions for studies involving metal-based catalysts, biochemical reactions, and materials science.
The chemical and physical properties of ammonium nickel sulfate make it a valuable tool for researchers across multiple disciplines.
Related compounds like 3,3′-diaminobenzidine, a chromogenic substrate used in immunohistochemistry, and the Vectastain Elite ABC kit, which utilizes the avidin-biotin complex (ABC) to amplify signal detection, are often used in conjunction with ammonium nickel sulfate in various experimental setups.
Salts such as NaCl, KCl, and sodium hydroxide may also be employed in the preparation and analysis of ammonium nickel sulfate samples.
Ammonium nickel sulfate's versatility and its role as a source of nickel ions make it a key component in a wide range of research applications, from catalysis and materials science to biochemistry and cell biology.
Optimizing the use of ammonium nickel sulfate and related compounds can be greatly facilitated by AI-driven research platforms like PubCompare.ai, which help researchers identify the most effective and reproducible protocols and products for their specific needs.
This green crystalline solid is soluble in water and has a wide range of industrial and research applications.
As a metal salt, ammonium nickel sulfate is used in electroplating, pigments, and as a precursor for other nickel compounds.
In scientific research, it is often employed as a source of nickel ions for studies involving metal-based catalysts, biochemical reactions, and materials science.
The chemical and physical properties of ammonium nickel sulfate make it a valuable tool for researchers across multiple disciplines.
Related compounds like 3,3′-diaminobenzidine, a chromogenic substrate used in immunohistochemistry, and the Vectastain Elite ABC kit, which utilizes the avidin-biotin complex (ABC) to amplify signal detection, are often used in conjunction with ammonium nickel sulfate in various experimental setups.
Salts such as NaCl, KCl, and sodium hydroxide may also be employed in the preparation and analysis of ammonium nickel sulfate samples.
Ammonium nickel sulfate's versatility and its role as a source of nickel ions make it a key component in a wide range of research applications, from catalysis and materials science to biochemistry and cell biology.
Optimizing the use of ammonium nickel sulfate and related compounds can be greatly facilitated by AI-driven research platforms like PubCompare.ai, which help researchers identify the most effective and reproducible protocols and products for their specific needs.