Human brain tissues from four sporadic AD patients, three Down syndrome patients with abundant tau pathology qualified for AD (referred to as AD/DS), and two normal controls were used in this study (Table S1). All cases used were histologically confirmed. Two of the AD/DS cases were provided by the University of Washington brain bank. The use of postmortem brain tissues for research was approved by the University of Pennsylvania’s Institutional Review Board with informed consent from patients or their families. For each purification, 6–14 g of frontal cortical gray matter was homogenized using a Dounce homogenizer in nine volumes (v/w) of high-salt buffer (10 mM Tris-HCl, pH 7.4, 0.8 M NaCl, 1 mM EDTA, and 2 mM dithiothreitol [DTT], with protease inhibitor cocktail, phosphatase inhibitor, and PMSF) with 0.1% sarkosyl and 10% sucrose added and centrifuged at 10,000 g for 10 min at 4°C. Pellets were reextracted once or twice using the same buffer conditions as the starting materials, and the supernatants from all two to three initial extractions were filtered and pooled. Additional sarkosyl was added to the pooled low-speed supernatant to reach 1%. After 1-h nutation at room temperature, samples were centrifuged again at 300,000 g for 60 min at 4°C. The resulted 1% sarkosyl-insoluble pellets, which contain pathological tau, were washed once in PBS and then resuspended in PBS (∼100 µl/g gray matter) by passing through 27-G 0.5-in. needles. The resuspended sarkosyl-insoluble pellets were further purified by a brief sonication (20 pulses at ∼0.5 s/pulse) using a hand-held probe (QSonica) followed by centrifugation at 100,000 g for 30 min at 4°C, whereby the majority of protein contaminants were partitioned into the supernatant, with 60–70% of tau remaining in the pellet fraction. The pellets were resuspended in PBS at one fifth to one half of the precentrifugation volume, sonicated with 20–60 short pulses (∼0.5 s/pulse), and spun at 10,000 g for 30 min at 4°C to remove large debris. The final supernatants, which contained enriched AD PHFs, were used in the study and referred to as AD-tau. In a subset of the experiments, the samples were boiled for 10 min right before the final 10,000-g spin to get rid of contaminating protease activity. The same purification protocol was used to prepare brain extracts from the two normal controls. The different fractions from PHF purification were characterized by Ponceau S staining, Western blotting (refer to Table S3 for antibodies), and sandwich ELISA for tau. The final supernatant fraction was further analyzed by transmission EM, BCA assay (Thermo Fisher Scientific), silver staining (SilverQuest Silver Staining kit; Thermo Fisher Scientific), and sandwich ELISA for Aβ 1–40, Aβ 1–42, and α-syn. The frontal cortex from one AD/DS case was purified using the traditional procedure with sucrose gradient fractionation as previously reported (Boluda et al., 2015 (link)). Enriched AD PHFs prepared using both methods showed similar seeding activity in primary hippocampal neurons from CD1 (non-Tg) mice.
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Ponceau S
Ponceau S
Ponceau S is a red dye used in biochemical and molecular biology applications, particularly for the staining of proteins in Western blot and SDS-PAGE analyses.
It provides a rapid, reversible method for visualizing protein bands on membranes or gels, allowing for easy identification and quantification.
Ponceau S staining is a simple, cost-effective technique that can be used to assess protein transfer efficiency and normalize protein loading prior to immunoblotting.
Researchers utilziing Ponceau S should be aware of its limitations, as it may not detect low-abundance proteins and can interfere with some downstream applications if not properly removed.
It provides a rapid, reversible method for visualizing protein bands on membranes or gels, allowing for easy identification and quantification.
Ponceau S staining is a simple, cost-effective technique that can be used to assess protein transfer efficiency and normalize protein loading prior to immunoblotting.
Researchers utilziing Ponceau S should be aware of its limitations, as it may not detect low-abundance proteins and can interfere with some downstream applications if not properly removed.
Most cited protocols related to «Ponceau S»
Antibodies
ARID1A protein, human
Autopsy
Biological Assay
Brain
Buffers
Centrifugation
Cortex, Cerebral
Dithiothreitol
Down Syndrome
Edetic Acid
Enzyme-Linked Immunosorbent Assay
Ethics Committees, Research
Fractionation, Chemical
Gray Matter
Homo sapiens
Lobe, Frontal
Mice, Laboratory
Needles
Neurons
Patients
Pellets, Drug
Peptide Hydrolases
Phosphoric Monoester Hydrolases
ponceau S
Protease Inhibitors
Proteins
Pulse Rate
Pulses
Sodium Chloride
sodium lauroyl sarcosinate
Sucrose
Tissues
Transmission, Communicable Disease
Tromethamine
For Figure 1 , our procedure for obtaining cytoplasmic and nuclear protein fractions has been described previously (29 (link)). To obtain the insoluble protein fraction, the pellet that was not soluble in either the hypertonic or nuclear lysis buffers was resuspended in the same volume of cell lysis buffer as used to obtain soluble protein fractions (550 μl). The 10% Bis–Tris NuPage Gel (Invitrogen) was loaded with 20 μg of soluble protein and 40 μl of insoluble suspension, transferred via iBLOT (Invitrogen) to a nitrocellulose membrane, immunoblotted for HA and β-actin, and imaged as previously described (29 (link)) except that rat α-HA (3F10, Roche) diluted 1:1000 and goat anti-rat 800 diluted 1:15 000 were used to detect HA. For Figure 7 , cells were harvested by scraping in PBS, washed once in PBS, and frozen. RIPA (Sigma) supplemented with PhosSTOP (Roche) and cOmplete ULTRA Tablets (Roche) inhibitors was added to each thawed cell pellet and cells were further lysed by freeze-thaw. Protein lysates were analyzed by Pierce BCA Protein Assay Kit (Thermo Scientific) and 25 μg protein was combined with NuPAGE sample buffer and reducing agent (Life Technologies), loaded onto 4–12% Bis–Tris gels (Life Technologies), and processed further as described above. Image Studio software (LiCOR) was used for quantification and further analysis was performed in Excel and GraphPad Prism.
For detection of Flag, the buffers and washes were Tris and milk-based as described elsewhere (30 (link)). The amount of unpurified protein sample loaded was 30 μl while the amount of purified protein sample loaded was 20 μl. The primary rabbit α-Flag antibody was diluted 1:1000 (Cell Signalling, Danvers, MA) and the secondary antibody was goat anti-rabbit HRP diluted 1:10 000 (Bio-Rad, Hercules, CA). The membrane was developed with SuperSignal West Pico Chemiluminescent Substrate (Thermo Pierce) and imaged on a ChemiDoc XRS+ (Bio-Rad). The membrane was stained with Ponceau S Staining Solution (Tocris Biosciences, Bristol, UK).
For detection of Flag, the buffers and washes were Tris and milk-based as described elsewhere (30 (link)). The amount of unpurified protein sample loaded was 30 μl while the amount of purified protein sample loaded was 20 μl. The primary rabbit α-Flag antibody was diluted 1:1000 (Cell Signalling, Danvers, MA) and the secondary antibody was goat anti-rabbit HRP diluted 1:10 000 (Bio-Rad, Hercules, CA). The membrane was developed with SuperSignal West Pico Chemiluminescent Substrate (Thermo Pierce) and imaged on a ChemiDoc XRS+ (Bio-Rad). The membrane was stained with Ponceau S Staining Solution (Tocris Biosciences, Bristol, UK).
Actins
Biological Assay
Bistris
Buffers
Cells
Cytoplasm
Freezing
Gels
Goat
Immunoglobulins
inhibitors
Milk, Cow's
Nitrocellulose
Nuclear Protein
ponceau S
prisma
Proteins
Rabbits
Radioimmunoprecipitation Assay
Reducing Agents
Tissue, Membrane
Tromethamine
The samples were dissolved in 2 μl 2× SDS-sample buffer per embryo and incubated for 5 min at 95°C. No homogenisation was necessary since the cells dissolved rapidly in the buffer. After full speed centrifugation for 1 min in a microcentrifuge to remove insoluble particles, samples were loaded on a gel (per lane 10–15 embryos for a minigel, 15–30 embryos for large format gels). If not enough sample buffer was added, a slurry formed (presumably from DNA). If a semi-quantitative analysis is needed, care should be taken to load the same amounts of embryos in every lane. Since the volume of cells and remaining supernatant after deyolking is hard to control, we suggest to (1) either deyolk only the desired number of embryos and to load the whole sample or to (2) deyolk a sufficiently large number of embryos so that the volume of cells and remaining supernatant can be neglected against the large volume of added sample buffer.
Electrophoresis, blotting and detection was performed essentially as described in [13 ]. Briefly, 10% SDS-gels (10 × 10 cm) were run, semi-dry-blotted onto PVDF-membrane as described in the manual (Immobilon P, Millipore), stained for 5 min with Ponceau S, blocked 1 h in 5% nonfat dry milk in PBST (0.5% Tween-20 in PBS), incubated over night at 4°C with primary antibody in blocking buffer, washed 30 min with 4 changes of PBST, incubated 1 h with secondary antibody in blocking buffer and washed 30 min with 4 changes of PBST. Membranes were incubated for 5 min in ECL Plus (Amersham Biosciences) and emitted light was detected using a cooled CCD-camera (LAS-1000, Fujifilm).
Electrophoresis, blotting and detection was performed essentially as described in [13 ]. Briefly, 10% SDS-gels (10 × 10 cm) were run, semi-dry-blotted onto PVDF-membrane as described in the manual (Immobilon P, Millipore), stained for 5 min with Ponceau S, blocked 1 h in 5% nonfat dry milk in PBST (0.5% Tween-20 in PBS), incubated over night at 4°C with primary antibody in blocking buffer, washed 30 min with 4 changes of PBST, incubated 1 h with secondary antibody in blocking buffer and washed 30 min with 4 changes of PBST. Membranes were incubated for 5 min in ECL Plus (Amersham Biosciences) and emitted light was detected using a cooled CCD-camera (LAS-1000, Fujifilm).
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Antibodies, Blocking
Cardiac Arrest
Cells
Centrifugation
Electrophoresis
Embryo
Gels
Immobilon P
Immunoglobulins
Light
Milk, Cow's
polyvinylidene fluoride
ponceau S
Tissue, Membrane
Tween 20
anti-IgG
Antibodies
Clone Cells
Goat
Immunoglobulins
Mice, House
Milk, Cow's
Peroxidase
Rabbits
Tissue, Membrane
For streptavidin blotting, HEK293T cells in wells of a 6-well plate were transfected with APXNES variants and labeled under the same conditions described for biotin-phenol imaging above. After 1 minute of labeling, the cells were washed 3 times with quencher solution (10 mM sodium azide, 10 mM sodium ascorbate, and 5 mM Trolox), then scraped and pelleted by centrifugation at 200 rpm for 5 minutes. The pellet was stored at -80 °C, then lysed with RIPA lysis buffer (50 mM Tris, 150 mM NaCl, 0.1% SDS, 0.5% sodium deoxycholate, 1% Triton X-100, 1 × protease cocktail (Sigma Aldrich, catalog no. P8849), 1 mM PMSF (phenylmethylsulfonyl fluoride), 10 mM sodium azide, 10 mM sodium ascorbate, and 5 mM Trolox) for 1 minute at 4 °C. The cell pellet was resuspended by gentle pipetting. Lysates were clarified by centrifugation at 13,000 rpm for 10 minutes at 4 °C before separation on a 9% SDS-PAGE gel. For blotting analysis, gels were transferred to nitrocellulose membrane, stained by Ponceau S (10 minutes in 0.1% w/v Ponceau S in 5% acetic acid/water), and blocked with “blot blocking buffer” (3% w/v BSA and 0.1% Tween-20 in Tris-buffered saline) at 4 °C overnight. The blots were immersed in streptavidin-HRP in blot blocking buffer (1:3000 dilution, Thermo Scientific) at room temperature for 60 minutes, then rinsed with blot blocking buffer 5 × 5 minutes before development with Clarity™ reagent (Bio-Rad) and imaging on an Alpha Innotech gel imaging system. For assessing comparative enzyme expression level, an identical gel and blot was prepared in parallel, and immunoblotted with α-FLAG-M2-HRP (1:3000, Sigma).
For blotting of endogenous proteins, HEK293T cells were grown in 6-well plates and infected at ∼50% confluency with 500 μL lentivirus prepared as described above. After 48 hours, cells were split into T25 flasks. From the same suspensions, cells were also plated into 48-well plates for side-by-side biotinphenol and immunofluorescence to check for enzyme activity and comparable enzyme expression levels. Biotin phenol labeling in the T25 flasks was performed with 30 minutes preincubation of 500 μM biotinphenol in 4 mL of a 1:1 mixture of DMEM:MEM (both from Cellgro) at 37°C. Flasks were quickly inverted so biotin-phenol media was on the ceiling of the flask, and 40 μL of 100 mM H2O2 was added (for a final concentration of 1 mM) to the media and agitated to mix. Flasks were again inverted and cells were exposed to the 1 mM H2O2 and 500 μM biotin-phenol solution for 1 minute at room temperature. After the 1 minute duration, the flask was inverted and the solution replaced with 5 mL of ice cold DPBS with quenchers (5 mM Trolox, 10 mM sodium ascorbate, 10 mM sodium azide) and the flask re-inverted for 1 minute. The flask was then washed with 5 mL DPBS + quenchers two more times. Cells were then resuspended by electronic pipette in 5 mL DPBS + quenchers and pelleted at 3000g for 10 minutes at 4°C. The supernatant was removed and the pellet was frozen at -80°C overnight.
Enrichment of endogenous biotinylated proteins was performed according to previous protocols. Cell pellets were lysed in 600 μL RIPA buffer (50 mM Tris, 150 mM NaCl, 0.1% SDS, 0.5% sodium deoxycholate, 1% Triton X-100, pH 7.5), with 1 × protease cocktail (Sigma Aldrich, catalog no. P8849), 1 mM PMSF (phenylmethylsulfonyl fluoride), 5 mM Trolox, 10 mM sodium ascorbate, and 10 mM sodium azide. The lysates were centrifuged at 15,000g for 10 minutes at 4°C and the supernatant was transferred to a new 1.5 mL microcentrifuge tube as whole cell lysate (WCL). Protein concentrations of each WCL were determined by the Pierce 660 nm Protein Assay (catalog no. 22660).
Streptavidin-coated magnetic beads (Pierce, catalog no. 88817) were washed twice with RIPA buffer and 1750 μg of each WCL sample was incubated with 120 μL of magnetic bead slurry in separate 1.5 mL microcentrifuge tubes with rotation for 1 hour at room temperature. The beads were subsequently washed twice with 1 mL RIPA lysis buffer, once with 1 mL of 1 M KCl in ddH2O, once with 1 mL of 0.1M Na2CO3 in ddH2O, once with 1 mL of 2 M urea in 10 mM Tris-HCl pH 8.0, and twice with 1 mL RIPA lysis buffer. Biotinylated proteins were eluted by boiling the beads in 75 μl 3 × protein loading buffer supplemented with 20 mM dithiothreitol (DTT) and 2 mM free biotin. 17.5 μg of each WCL and 25 μl of each streptavidin eluate (SAE) were separated on 9% SDS-PAGE gels.
Nitrocellulose blot transfer and Ponceau S staining were performed as described above. Membranes were blocked with blot blocking buffer overnight at 4°C, cut into strips based on MW, and incubated facedown into 500 μl of blot blocking buffer containing the specified primary antibodies (see table below) for 1 hour at room temperature. Blots were washed 5 × 5 minutes in blot blocking buffer before incubation in 10 mL of secondary antibody in blot blocking buffer for 1 hour at room temperature. The membranes were then washed again 5 × 5 minutes in blot blocking buffer before imaging with Clarity™ reagent (Bio-Rad) as described above.
For blotting of endogenous proteins, HEK293T cells were grown in 6-well plates and infected at ∼50% confluency with 500 μL lentivirus prepared as described above. After 48 hours, cells were split into T25 flasks. From the same suspensions, cells were also plated into 48-well plates for side-by-side biotinphenol and immunofluorescence to check for enzyme activity and comparable enzyme expression levels. Biotin phenol labeling in the T25 flasks was performed with 30 minutes preincubation of 500 μM biotinphenol in 4 mL of a 1:1 mixture of DMEM:MEM (both from Cellgro) at 37°C. Flasks were quickly inverted so biotin-phenol media was on the ceiling of the flask, and 40 μL of 100 mM H2O2 was added (for a final concentration of 1 mM) to the media and agitated to mix. Flasks were again inverted and cells were exposed to the 1 mM H2O2 and 500 μM biotin-phenol solution for 1 minute at room temperature. After the 1 minute duration, the flask was inverted and the solution replaced with 5 mL of ice cold DPBS with quenchers (5 mM Trolox, 10 mM sodium ascorbate, 10 mM sodium azide) and the flask re-inverted for 1 minute. The flask was then washed with 5 mL DPBS + quenchers two more times. Cells were then resuspended by electronic pipette in 5 mL DPBS + quenchers and pelleted at 3000g for 10 minutes at 4°C. The supernatant was removed and the pellet was frozen at -80°C overnight.
Enrichment of endogenous biotinylated proteins was performed according to previous protocols. Cell pellets were lysed in 600 μL RIPA buffer (50 mM Tris, 150 mM NaCl, 0.1% SDS, 0.5% sodium deoxycholate, 1% Triton X-100, pH 7.5), with 1 × protease cocktail (Sigma Aldrich, catalog no. P8849), 1 mM PMSF (phenylmethylsulfonyl fluoride), 5 mM Trolox, 10 mM sodium ascorbate, and 10 mM sodium azide. The lysates were centrifuged at 15,000g for 10 minutes at 4°C and the supernatant was transferred to a new 1.5 mL microcentrifuge tube as whole cell lysate (WCL). Protein concentrations of each WCL were determined by the Pierce 660 nm Protein Assay (catalog no. 22660).
Streptavidin-coated magnetic beads (Pierce, catalog no. 88817) were washed twice with RIPA buffer and 1750 μg of each WCL sample was incubated with 120 μL of magnetic bead slurry in separate 1.5 mL microcentrifuge tubes with rotation for 1 hour at room temperature. The beads were subsequently washed twice with 1 mL RIPA lysis buffer, once with 1 mL of 1 M KCl in ddH2O, once with 1 mL of 0.1M Na2CO3 in ddH2O, once with 1 mL of 2 M urea in 10 mM Tris-HCl pH 8.0, and twice with 1 mL RIPA lysis buffer. Biotinylated proteins were eluted by boiling the beads in 75 μl 3 × protein loading buffer supplemented with 20 mM dithiothreitol (DTT) and 2 mM free biotin. 17.5 μg of each WCL and 25 μl of each streptavidin eluate (SAE) were separated on 9% SDS-PAGE gels.
Nitrocellulose blot transfer and Ponceau S staining were performed as described above. Membranes were blocked with blot blocking buffer overnight at 4°C, cut into strips based on MW, and incubated facedown into 500 μl of blot blocking buffer containing the specified primary antibodies (see table below) for 1 hour at room temperature. Blots were washed 5 × 5 minutes in blot blocking buffer before incubation in 10 mL of secondary antibody in blot blocking buffer for 1 hour at room temperature. The membranes were then washed again 5 × 5 minutes in blot blocking buffer before imaging with Clarity™ reagent (Bio-Rad) as described above.
Most recents protocols related to «Ponceau S»
Ponceau S solution was purchased from Sigma-Aldrich. Nanoparticle preparations were stained with 1 mL of Ponceau S solution for 2 min and then washed twice with distilled water to remove background stain by centrifuging at 15,000 × g for 5 min at 4 °C.
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Where indicated, gels after native PAGE or SDS-PAGE were washed with deionized water three times for 5 min and incubated with Coomassie G-250 stain (Bio-Rad) for 1 h. The gels were washed with water after to remove the excess of the dye and imaged. Where indicated, membranes after protein transfer were incubated with Ponceau S solution (Sigma) for 10 min, then were washed with water to remove the excess of the dye and imaged.
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Equal amounts of 200 µg of kidney cortical proteins were loaded per lane onto a polyacrylamide gel and subjected to electrophoresis. Following separation, the proteins were transferred onto nitrocellulose membranes. Membranes underwent treatment with a 0.1% Ponceau S solution (Sigma-Aldrich, St. Louis, MO, USA) for 10 min on a shaker, which was followed by rinsing with distilled water to eliminate background staining. Ponceau S staining served for total protein normalization.
The transferred proteins were then probed using specific antibodies, including a mouse eNOS antibody (1:250; BD610297BD, Biosciences, San Jose, CA, USA), a mouse nNOS antibody (1:200; SC-5302, Santa Cruz, CA, USA), a mouse DDAH1 antibody (1:500; SC-271337, Santa Cruz), or a rabbit DDAH2 antibody (1:2000; Ab184166, Abcam, Cambridge, UK). Subsequent to washing, the blots were incubated with the corresponding secondary antibody conjugated to horseradish peroxidase. Immunopositive bands were scanned using an imaging densitometer (Quantity One, Bio-Rad, Hercules, CA, USA) to quantify integrated optical density (IOD). Protein abundance was expressed as IOD normalized by Ponceau S stain (PonS, representing the total protein loaded). Complete blots and corresponding images of Ponceau S staining can be found inSupplementary Material .
The transferred proteins were then probed using specific antibodies, including a mouse eNOS antibody (1:250; BD610297BD, Biosciences, San Jose, CA, USA), a mouse nNOS antibody (1:200; SC-5302, Santa Cruz, CA, USA), a mouse DDAH1 antibody (1:500; SC-271337, Santa Cruz), or a rabbit DDAH2 antibody (1:2000; Ab184166, Abcam, Cambridge, UK). Subsequent to washing, the blots were incubated with the corresponding secondary antibody conjugated to horseradish peroxidase. Immunopositive bands were scanned using an imaging densitometer (Quantity One, Bio-Rad, Hercules, CA, USA) to quantify integrated optical density (IOD). Protein abundance was expressed as IOD normalized by Ponceau S stain (PonS, representing the total protein loaded). Complete blots and corresponding images of Ponceau S staining can be found in
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Firstly, in Supplementary Note S6 , we explored the linear relationship between the concentration of Ponceau S and its absorbance (Supplementary Fig. S11a, b ). During the test, the experimental group placed a capsule containing a known mass (m1) of Ponceau S (3 mg/ml) in a water tank containing a known mass (m2) of water. The capsule was fixed on a shaking table set at parameters of 100 rpm for 5 min. Simultaneously, a control group was established: the solution of Ponceau S with mass m1 was directly poured into the water with mass m2 and thoroughly stirred, representing 100% drug leakage. Each case was repeated three times, and the absorbance at the corresponding wavelengths of both the experimental control groups was measured using a UV-vis spectrometer (UV-2600i, SHIMADZU Co., Ltd.). The ratio of the absorbance of the experimental group to that of the control group can be considered as the proportion of the leaked capsule drug.
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We performed chemiluminescence capturing with automatic or manual exposure time as described in the Operating Instructions of the Amersham™ Imager 600 (GE Healthcare). A short pre-exposure is performed to determine the signal intensity. The system will use this information to calculate which exposure time will give the highest possible signal below saturation to enable accurate quantification of the sample.’ Manual exposure is only performed with a maximum duration of 5 min, in case the intensity of the image is inadequate (i.e., takes > 5–10 min) after automatic exposure, which is also suggested by the manufacturer’ instructions. Western blot images displayed in the (supplementary) figures of this manuscript have been adjusted for brightness and contrast equally throughout the picture. In addition, the Ponceau S band (37, 50, 70 or 75 kDa) presented in the (supplementary) figures is representative for the total Ponceau S staining per sample. For illustration purposes of the protein abundance in the human peripheral lung tissue homogenates in the manuscript figures, examples of bands of the target proteins along with the corresponding normalization bands of the Ponceau S staining are shown of one patient/group, which is not always representative for the mean and changes in the whole patient group as quantified in the box plots (due to the variation between patients). In the Supplementary Figs.1 –3 , all western blot images of the target proteins along with the corresponding normalization bands of the Ponceau S staining of all subjects/group are shown of the peripheral lung tissue from non-COPD patients and COPD patients (GOLDII; GOLDIV) (single) (Figure S1 ), as well as images of undifferentiated PBEC (pool of quintuplicates) (Figure S2 ) and differentiated PBEC (duplicates) (Figure S3 ) from non-COPD patients and COPD patients.
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Top products related to «Ponceau S»
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Ponceau S is a sensitive stain used in biochemistry and molecular biology to detect and quantify proteins on membranes, such as nitrocellulose or PVDF, after electrophoretic transfer or dot blotting. It reversibly binds to basic amino acid residues, allowing rapid visualization of protein bands or spots.
Sourced in United States, Germany, United Kingdom
Ponceau S solution is a staining reagent used in biochemistry and molecular biology laboratories. It is a water-soluble dye that binds to proteins, allowing for the visualization of protein bands on membranes, such as those used in Western blotting procedures.
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Nitrocellulose membranes are a type of laboratory equipment designed for use in protein detection and analysis techniques. These membranes serve as a support matrix for the immobilization of proteins, enabling various downstream applications such as Western blotting, dot blotting, and immunodetection.
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PVDF membranes are a type of laboratory equipment used for a variety of applications. They are made from polyvinylidene fluoride (PVDF), a durable and chemically resistant material. PVDF membranes are known for their high mechanical strength, thermal stability, and resistance to a wide range of chemicals. They are commonly used in various filtration, separation, and analysis processes in scientific and research settings.
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Image Lab software is a data analysis tool designed for use with Bio-Rad's gel and blot imaging systems. The software provides a user-friendly interface for capturing, analyzing, and processing images of gels, blots, and other samples.
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The Protease Inhibitor Cocktail is a laboratory product designed to inhibit the activity of proteases, which are enzymes that can degrade proteins. It is a combination of various chemical compounds that work to prevent the breakdown of proteins in biological samples, allowing for more accurate analysis and preservation of protein integrity.
Sourced in United Kingdom, Germany, United States
Ponceau S is a protein staining dye used for the detection and visualization of proteins on nitrocellulose or PVDF membranes after gel electrophoresis and Western blotting. It reversibly stains proteins with a pink-red color, allowing for the identification and quantification of protein bands.
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Nitrocellulose membranes are porous sheets made from nitrocellulose, a form of cellulose nitrate. They are commonly used in various laboratory techniques, such as Western blotting, immunodetection, and nucleic acid transfer, to immobilize and detect specific proteins, DNA, or RNA molecules.
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The Trans-Blot Turbo Transfer System is a laboratory instrument designed for efficient and rapid protein transfer from polyacrylamide gels to membranes. It utilizes a unique power supply and transfer cassette design to enable fast and consistent protein transfer, optimizing the time required for Western blotting procedures.
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Nitrocellulose membranes are a type of laboratory equipment used for the transfer and immobilization of proteins and nucleic acids. They are commonly utilized in various analytical techniques, such as Western blotting and dot blotting, to facilitate the detection and identification of specific biomolecules.
More about "Ponceau S"
Ponceau S, also known as Acid Red 112, is a widely used protein stain in biochemistry and molecular biology.
It is a red dye that binds to basic amino acids, allowing for the rapid, reversible visualization of protein bands on membranes or gels, such as those used in Western blot and SDS-PAGE analyses.
This simple and cost-effective staining technique is particularly useful for assessing protein transfer efficiency and normalizing protein loading prior to immunoblotting.
Ponceau S staining is a valuable tool for researchers, as it provides a quick and easy way to confirm the presence and relative abundance of proteins of interest.
It can be used on nitrocellulose or PVDF membranes, and the stained proteins can be easily quantified using image analysis software like Image Lab.
While Ponceau S is a versatile stain, it's important to be aware of its limitations.
It may not detect low-abundance proteins, and the stain can interfere with some downstream applications if not properly removed.
To mitigate these issues, researchers often incorporate a protease inhibitor cocktail into their Ponceau S staining protocols.
By leveraging the insights gained from the MeSH term description and the Metadescription, researchers can optimize their Ponceau S protocols and improve the reproducibility and accuracy of their protein analysis workflows.
PubCompare.ai's AI-driven research protocol comparisons can be a valuable resource in this process, helping researchers identify the most effective Ponceau S staining methods from the literature, preprints, and patents.
It is a red dye that binds to basic amino acids, allowing for the rapid, reversible visualization of protein bands on membranes or gels, such as those used in Western blot and SDS-PAGE analyses.
This simple and cost-effective staining technique is particularly useful for assessing protein transfer efficiency and normalizing protein loading prior to immunoblotting.
Ponceau S staining is a valuable tool for researchers, as it provides a quick and easy way to confirm the presence and relative abundance of proteins of interest.
It can be used on nitrocellulose or PVDF membranes, and the stained proteins can be easily quantified using image analysis software like Image Lab.
While Ponceau S is a versatile stain, it's important to be aware of its limitations.
It may not detect low-abundance proteins, and the stain can interfere with some downstream applications if not properly removed.
To mitigate these issues, researchers often incorporate a protease inhibitor cocktail into their Ponceau S staining protocols.
By leveraging the insights gained from the MeSH term description and the Metadescription, researchers can optimize their Ponceau S protocols and improve the reproducibility and accuracy of their protein analysis workflows.
PubCompare.ai's AI-driven research protocol comparisons can be a valuable resource in this process, helping researchers identify the most effective Ponceau S staining methods from the literature, preprints, and patents.