Log In Sign up
The largest database of trusted experimental protocols
> Chemicals & Drugs > Organic Chemical > Nitrocellulose

Nitrocellulose

Nitrocellulose is a highly flammable compound derived from the nitration of cellulose.
It has a variety of applications, including in lacquers, explosives, and as a binder in paints and photographic films.
Nitrocellulose research requires careful planning and execution to ensure reproducibility and accuracy.
PubCompare.ai, the leading AI-driven platform, can help optimize your nitrocellulose research by easily locating the best protocols from literature, pre-prints, and patents using advanced AI-powered comparisons.
Discover the most reliable and effective nitrocellulose-related products and methodologies to elevate your research outcomes.
Experiennce the power of PubCompare.ai today and take your nitrocellulose research to new heights.

Most cited protocols related to «Nitrocellulose»

1
Intracellular Labeling of Cortical Neurons
Four 9 month old male mice (C57Bl/SJL) were used. Animals were anesthetized with choral hydrate (15% aqueous solution, i.p.) and were perfused transcardially with 4% paraformaldehyde and 0.125% glutaraldehyde in phosphate buffer saline (PBS; pH 7.4). The brains were then carefully removed from the skull and postfixed for 6 hours. All procedures were conducted in accordance with the National Institute of Health Guide for the Care and Use of Laboratory Animals and were approved by the Mount Sinai School of Medicine Institutional Animal Care and Uses Committee.
For intracellular injections, brains were coronally sectioned at 200 µm on a Vibratome (Leica, Nussloch, Germany). The sections were then incubated in 4,6-diamidino-2-phenylindole (DAPI; Sigma, St. Louis, MO, USA), a fluorescent nucleic acid stain, for 5 minutes, mounted on nitrocellulose filter paper and immersed in PBS. Using DAPI as a staining guide, individual layer II/III pyramidal neurons of the frontal cortex were loaded with 5% Lucifer Yellow (Molecular Probes, Eugene, OR, USA) in distilled water under a DC current of 3–8 nA for 10 minutes, or until the dye had filled distal processes and no further loading was observed [45] (link), [49] (link). Tissue slices were then mounted and coverslipped in Permafluor. Dendritic segment and spine imaging was performed using a Zeiss 410 confocal laser scanning microscope (Zeiss, Thornwood, NY, USA) using a 488 nm excitation wavelength, using a 1.4 N.A. Plan-Apochromat 100× objective with a working distance of 170 µm and a 5× digital zoom. After gain and offset settings were optimized, segments were digitally imaged at 0.1 µm increments, along the optical axis. The confocal stacks were then deconvolved with AutoDeblur (MediaCybernetics, Bethesda, MD, USA).
Supporting Information is available online (Box S1)
Rodriguez A., Ehlenberger D.B., Dickstein D.L., Hof P.R, & Wearne S.L. (2008). Automated Three-Dimensional Detection and Shape Classification of Dendritic Spines from Fluorescence Microscopy Images. PLoS ONE, 3(4), e1997.
Full text: Click here
Publication 2008
Animals Animals, Laboratory Brain Buffers Cranium DAPI Dendrites Epistropheus Fingers Glutaral Lobe, Frontal lucifer yellow Males Mice, House Microscopy, Confocal Molecular Probes Nitrocellulose Nucleic Acids paraform Phosphates Protoplasm Pyramidal Cells Saline Solution Stains Tissues Vertebral Column Vision
2
Detailed eCLIP Experiment Protocol
(See Supplementary Protocol 1 for detailed Standard Operating Procedures for ENCODE-style eCLIP experiments, including oligonucleotide sequences, catalog numbers for all reagents, and specific details for eCLIP experiments). RNA binding protein (RBP)-RNA interactions were stabilized with UV crosslinking (254 nm, 400 mJ/cm2), followed by lysis in iCLIP lysis buffer, limited digestion with RNase I (Ambion), immunoprecipitation of RBP-RNA complexes with a specific primary antibody of interest using magnetic beads with pre-coupled secondary antibody (typically M-280 Sheep Anti-Rabbit IgG Dynabeads, ThermoFisher Scientific 11204D), and stringent washes. After dephosphorylation with FastAP (ThermoFisher) and T4 PNK (NEB), a barcoded RNA adapter was ligated to the 3′ end (T4 RNA Ligase, NEB) (at this step, multiple replicates of the same RBP, or potentially RBPs of similar size and bound RNA amount, can be uniquely barcoded and pooled after ligation to simplify downstream steps - see Supplementary Fig. 2a). Ligations were performed on-bead (to allow washing away unincorporated adapter) in high concentration of PEG8000, which improves ligation efficiency to > 90%. Samples were then run on standard protein gels and transferred to nitrocellulose membranes, and a region 75 kDa (~150 nt of RNA) above the protein size was isolated and proteinase K (NEB) treated to isolate RNA. RNA was reverse transcribed with AffinityScript (Agilent), and treated with ExoSAP-IT (Affymetrix) to remove excess oligonucleotides. A second DNA adapter (containing a random-mer of 5 (N5) or 10 (N10) random bases at the 5′ end) was then ligated to the cDNA fragment 3′ end (T4 RNA Ligase, NEB), performed with high concentration of PEG8000 (to improve ligation efficiency) and DMSO (to decrease inhibition of ligation due to secondary structure). After cleanup (Dynabeads MyOne Silane, ThermoFisher), an aliquot of each sample was first subjected to qPCR (to identify the proper number of PCR cycles), and then the remainder was PCR amplified (Q5, NEB) and size selected via agarose gel electrophoresis. Samples were sequenced on the Illumina HiSeq 2500 or 4000 platform as two Paired End 50bp (for N5) or 55bp (for N10) reads. All analyses were performed using identical antibody lots for RBFOX2 (A300-864A lot 002, Bethyl), SLBP (RN045P lot 001, MBL International), and IgG Isotype Control (02-6102 lot 32013, Thermo Fisher Scientific). SLBP experiments were performed with 20×106 cells and 10 ug of primary antibody; RBFOX2 experiments were performed with 20×106 cells and 10 ug (eCLIP Rep1 and Rep2) or 10×106 cells and 5 ug (RNase I variation experiments). All experiments in K562 and HepG2 cells were performed with 20×106 cells and 10 ug of indicated primary antibody (Supplementary Table 2). Antibody validation documentation (including Western images of immunoprecipitation and shRNA knockdown19 (link)) are available at http://www.encodeproject.org/. Additional experiments performed in K562 and HepG2 cells in which the antibody failed to successfully immunoprecipitate the targeted RBP were excluded from analysis. 293T cells were obtained from Clontech (Lenti-X 293T cell line). K562 and HepG2 cells were purchased from ATCC, and were not independently verified. Cells were routinely tested for mycoplasma using MycoAlert PLUS (Lonza).
Van Nostrand E.L., Pratt G.A., Shishkin A.A., Gelboin-Burkhart C., Fang M.Y., Sundararaman B., Blue S.M., Nguyen T.B., Surka C., Elkins K., Stanton R., Rigo F., Guttman M, & Yeo G.W. (2016). Robust transcriptome-wide discovery of RNA binding protein binding sites with enhanced CLIP (eCLIP). Nature methods, 13(6), 508-514.
Publication 2016
anti-IgG Buffers Cell Lines Cells Digestion DNA, Complementary Domestic Sheep Electrophoresis, Agar Gel Endopeptidase K Gels HEK293 Cells Hep G2 Cells Immunoglobulin Isotypes Immunoglobulins Immunoprecipitation Ligation M 280 Mycoplasma Nitrocellulose Oligonucleotides polyethylene glycol 8000 Proteins Psychological Inhibition Rabbits Ribonuclease, Pancreatic RNA-Binding Proteins RNA Ligase (ATP) Short Hairpin RNA Silanes Sulfoxide, Dimethyl Tissue, Membrane
3
Mosquito Embryo Microinjection and Transgenesis
A. gambiae mosquitoes were maintained in standard insectary conditions (28°, 75–80% humidity, 12-hr/12-hr light/dark cycle). Larvae were raised in deionized water and fed finely ground TetraMin fish food. Embryo microinjection was performed essentially as described (Fuchs et al. 2013 (link); Pondeville et al. 2014 (link)). Freshly laid eggs were directly aligned against the edge of a nitrocellulose membrane kept wet with overlaying filter paper soaked with demineralized water. A mix of plasmids totaling 400 ng/µl of DNA (0, 1 mM NaHPO4 buffer pH 6.8, 5 mM KCl, 60 ng/µl helper plasmid, and generally 85 ng/µl of each of four distinct transgenesis plasmids) was injected under a Nikon Eclipse TE2000-S inverted microscope using an Eppendorf Femtojet injector and TransferMan NK2 micromanipulator. Injections were performed using the compensation pressure of the device, which was kept at 6000 hPa to promote a constant moderate flow of the DNA solution out of the quartz capillary. Microinjected eggs were left undisturbed on the injection slides, which were placed diagonally in a container with 1-cm-deep demineralized water, the part of the filter paper most distant from the eggs was dipped in water so that eggs remained wet by capillarity (Figure 1). Adult mosquitoes that survived microinjection were separated according to sex and crossed en masse to an excess of fresh wild-type adults. Neonate progeny larvae from several successive gonotrophic cycles were screened by spotting groups of 50–80 onto the wells of a 24-well teflon-coated diagnostic slide (Erie Scientific, Menzel GmbH, Braunschweig, Germany) under a Zeiss Axiovert 200M fluorescence microscope. When a fluorescent larva was detected, it was carefully isolated from the remainder larvae with the cut tip of a P200 pipette.
Volohonsky G., Terenzi O., Soichot J., Naujoks D.A., Nolan T., Windbichler N., Kapps D., Smidler A.L., Vittu A., Costa G., Steinert S., Levashina E.A., Blandin S.A, & Marois E. (2015). Tools for Anopheles gambiae Transgenesis. G3: Genes|Genomes|Genetics, 5(6), 1151-1163.
Full text: Click here
Publication 2015
Adult Buffers Capillaries Capillarity Culicidae Diagnosis DNA, A-Form Eggs Embryo Fishes Food Humidity Infant, Newborn Larva Medical Devices Microinjections Microscopy Microscopy, Fluorescence Nitrocellulose Plasmids Pressure Quartz Teflon Tissue, Membrane
4
Western Blot Quantification of Cellular Signaling Proteins
The MCF-7 cell line was maintained under standard conditions in Dulbecco's modified Eagle's medium supplemented with 10% foetal bovine serum. Cells were washed with ice cold phosphate buffered Saline and lysed in RIPA buffer (1% NP-40, 0.1% SDS, 0.5% Sodium deoxycholate, 50 mM Tris pH 7.5, 150 mM NaCl) supplemented with protease and phosphatase inhibitor cocktails (Sigma Aldrich) and protein concentration was quantitated by BCA protein assay (Invitrogen). Purified BSA (Applichem) was dissolved in RIPA buffer. Cell lysates and a BSA sample were serially diluted 1∶2 and run on SDS-PAGE using a standard protocol. Proteins were transferred to the PVDF (for ECL based detection) or Nitrocellulose (for LI-COR based proteins detection) membranes. Membranes were blocked with blocking solution (11500694001, Roche) for BSA detection or 5% skimmed milk for rest of the membranes. For Western blotting ERK (M-5670, Sigma Aldrich), mTOR (2972, Cell Signaling Technology), RSK1 (sc-231, Santa Cruz) and BSA (sc-50528, Santa Cruz) antibodies were used. Anti-rabbit HRP-conjugated (Cell Signaling Technology) or anti-Rabbit IR 800 (LI-COR) secondary antibodies were used for ECL or LI-COR protein detection systems, respectively. Signal was detected by standard X-ray films (Fuji), CCD camera (Advanced Molecular Vision) or LI-COR scanner.
Degasperi A., Birtwistle M.R., Volinsky N., Rauch J., Kolch W, & Kholodenko B.N. (2014). Evaluating Strategies to Normalise Biological Replicates of Western Blot Data. PLoS ONE, 9(1), e87293.
Full text: Click here
Publication 2014
Antibodies Biological Assay Buffers Cells Cold Temperature Deoxycholic Acid, Monosodium Salt Fetal Bovine Serum FRAP1 protein, human MCF-7 Cells Milk, Cow's Nitrocellulose Nonidet P-40 Peptide Hydrolases Phosphates Phosphoric Monoester Hydrolases polyvinylidene fluoride Proteins Rabbits Radioimmunoprecipitation Assay RPS6KA1 protein, human Saline Solution SDS-PAGE Sodium Chloride Tissue, Membrane Tromethamine Vision X-Ray Film
5
SDS-PAGE and Western Blot Analysis
Protein samples were treated for 30 min at 50°C in a sample-treating solution containing 2% SDS and 5% β-mercaptoethanol. The samples were then loaded onto a 12% or 15% SDS-polyacrylamide gel (7.0×8.3 cm ×0.75 mm) and electrophoresed using Mini-Protean Tetra system (Bio-Rad) at 15 mA (current constant). After electrophoresis, proteins separated on the gel were transferred onto a methanol-activated PVDF membrane (Immobilon-P, pore size 0.45 µm, Millipore) or a nitrocellulose membrane (Protran BA85, pore size 0.45 µm, Whatman) for 2 h using TE22 Mighty Small Transfer system (Hoefer Scientific) at 100 V (voltage constant). The membrane was then treated with or without phosphate-buffered saline (PBS) containing 0.4% PFA for 30 min at room temperature, followed by blocking for 1 h with 5% skim milk (Carnation) in Tris-buffered saline containing 0.1% Tween-20 (TBS-T). The membrane was then incubated for 1 h with a primary antibody in TBS-T containing 1% skim milk. As primary antibody, mouse monoclonal anti-α-syn antibodies 4D6 and LB509 (Santa Cruz Biotechnology) and rabbit monoclonal anti-phospho α-syn antibody EP1536Y (Epitomics, Burlingame, CA) were used at a dilution 1∶1,000, and rabbit polyclonal anti-actin antibody (Sigma) was also used at a dilution 1∶5,000. After washing with TBS-T containing 1% skim milk for 5 min three times, the membrane was incubated for 1 h with a secondary antibody, horseradish peroxidase-conjugated anti-mouse IgG or anti-rabbit IgG antibody (Santa Cruz Biotechnology), in TBS-T containing 1% skim milk. After washing with TBS-T for 10 min three times, protein bands on the membrane were detected by chemiluminescence method using ECL-Plus immunoblotting detection system (GE Healthcare).
Lee B.R, & Kamitani T. (2011). Improved Immunodetection of Endogenous α-Synuclein. PLoS ONE, 6(8), e23939.
Full text: Click here
Publication 2011
2-Mercaptoethanol Actins Anti-Antibodies anti-IgG Antibodies, Anti-Idiotypic Carnation Chemiluminescence Electrophoresis Horseradish Peroxidase Immobilon P Immunoglobulins Membrane Proteins Methanol Mice, House Milk, Cow's Nitrocellulose Phosphates polyacrylamide gels polyvinylidene fluoride Proteins Rabbits Saline Solution Technique, Dilution Tetragonopterus Tissue, Membrane Tween 20

Most recents protocols related to «Nitrocellulose»

1
Potent PARG Inhibitor Development
Not available on PMC !

Example 2

Development of potent and cell-permeant PARG inhibitors via multi-step virtual screening and hierarchical selection. Forty candidates from National Cancer Institute (NCI) were selected to examine the efficacy of PARG inhibition by dot blot assay. PARG was incubated with PAR for 20 min at room temperature with or without inhibitors. PAR-digestion results were analyzed using dot blotting with anti-PAR antibody. Two compounds, #5 and #34, showed the good inhibitory activity for PARG. PC and NC mean positive control (PAR only) and negative control (no inhibitor), respectively. IC50 value of compound 34 was measured by dot blotting with anti-PAR antibody in a dose course of compound 34.

PAR digestion assay: Recombinant PARG protein were incubated with PAR (10 μM, calculated as the ADP-ribose unit) and DMSO (Negative control, NC) or small molecules for 20 minutes at room temperature. Positive control (PC) only contains PAR in PBS. Samples were spotted onto a nitrocellulose membrane. The membrane was blocked with TBST buffer (0.15 M NaCl, 0.01 M Tris-HCl at pH 7.4, 0.1% Tween 20) supplemented with 5% milk and extensively washed with TBST. The membrane was examined by anti-PAR antibody.

US11858879B2. PARG inhibitors and method of use thereof (2024-01-02). City of Hope [US]. Inventors: Xiaochun Yu [US], Shih-Hsun Chen [US], Yate-Ching Yuan [US], Hongzhi Li [US], David Horne [US].
Full text: Click here
Patent 2024
Adenosine Diphosphate Ribose Antibodies, Anti-Idiotypic Biological Assay Buffers Cells Digestion Dot Immunoblotting inhibitors Milk, Cow's Nitrocellulose Psychological Inhibition Recombinant Proteins Sodium Chloride Sulfoxide, Dimethyl Tissue, Membrane Tromethamine Tween 20
2
Quantitative Analysis of PAR Hydrolysis
Not available on PMC !

Example 9

Recombinant PAR was purified from a biochemical assay using PARP1. The concentration of PAR was calculated as the ADP-ribose unit. Recombinant full length PARG was incubated with 10 μM PAR in the presence of DMSO (negative control) or small molecule compounds (CHP20-25) in a 10 μl reaction for 20 minutes at room temperature. Positive control only contains PAR in PBS. For dot blotting analysis, samples (1 μl) were spotted onto a nitrocellulose membrane. Then, the membrane was baked for 30 minutes at 60° C. and blocked with TBST buffer (0.15 M NaCl, 0.01 M Tris-HCl at pH 7.4, 0.1% Tween 20) supplemented with 5% milk for 30 minutes at room temperature. After washing with TBST, the membrane was incubated with anti-PAR monoclonal antibody (Trevigen) overnight at 4° C. Following standard western blot method, the signals were visualized by chemiluminescent detection and results are shown in FIG. 7A-FIG. 7F.

US11858879B2. PARG inhibitors and method of use thereof (2024-01-02). City of Hope [US]. Inventors: Xiaochun Yu [US], Shih-Hsun Chen [US], Yate-Ching Yuan [US], Hongzhi Li [US], David Horne [US].
Full text: Click here
Patent 2024
Adenosine Diphosphate Ribose Biological Assay Buffers Digestion Dot Immunoblotting Milk, Cow's Monoclonal Antibodies Nitrocellulose PARP1 protein, human Sodium Chloride Sulfoxide, Dimethyl Tissue, Membrane Tromethamine Tween 20 Western Blotting
3
PARG-Mediated DePARylation Inhibition
Not available on PMC !

Example 6

Recombinant full length PARG protein is generated from Sf9 insect cells. Recombinant PAR is purified from a biochemical assay using PARP1. PARG is incubated with PAR in the presence of DMSO (Negative control, NC) or small chemical compounds for 20 minutes at room temperature. Positive control (PC) only contains PAR. Samples (1 μl) were spotted onto a nitrocellulose membrane. Then, the membrane was baked for 30 minutes at 60° C. and blocked with TBST buffer (0.15 M NaCl, 0.01 M Tris-HCl at pH 7.4, 0.1% Tween 20) supplemented with 5% milk for 30 minutes at room temperature. After washing with TBST, the membrane was incubated with monoclonal anti-PAR antibody (Trevigen, Inc.) for overnight at 4° C. Following standard western blot method, the signals were visualized by chemiluminescent detection. With the chemical inhibition of the dePARylation activity of PARG, we are able to detect the dot signals of PAR.

US11858879B2. PARG inhibitors and method of use thereof (2024-01-02). City of Hope [US]. Inventors: Xiaochun Yu [US], Shih-Hsun Chen [US], Yate-Ching Yuan [US], Hongzhi Li [US], David Horne [US].
Full text: Click here
Patent 2024
Antibodies, Anti-Idiotypic Biological Assay Buffers Chemical Actions compound 20 Digestion Insecta JTB protein, human Milk, Cow's Monoclonal Antibodies Nitrocellulose PARP1 protein, human Psychological Inhibition Sf9 Cells Sodium Chloride Sulfoxide, Dimethyl Tissue, Membrane Tromethamine Tween 20 Western Blotting
4
Sepsis Modeling in Mice Using N. meningitidis Strains
Not available on PMC !

Example 7

Sepsis modeling was performed as described by Gorringe A. R., Reddin, K. M., Voet P. and Poolman J. T. (Methods Mol. Med. 66, 241 (Jan. 1, 2001)) and Johswich, K. O. et al. (Infect. Immun. 80, 2346 (Jul. 1, 2012)). Groups of 6 eight-week-old C57BL/6 mice (Charles River Laboratories) were inoculated via intraperitoneal injection with N. meningitidis strain B16B6, B16B6 Δtbpb, or B16B6 Δnmb0313 (N=2 independent experiments). To prepare inoculums, bacterial strains for infection were grown overnight on GC agar, resuspended and then grown for 4 h in 10 ml of Brain Heart Infusion (BHI) medium at 37° C. with shaking. Cultures were adjusted such that each final 500 μl inoculum contained 1×106 colony forming units and 10 mg human holo-transferrin. Mice were monitored at least every 12 h starting 48 h before infection to 48 h after infection for changes in weight, clinical symptoms and bacteremia. Mice were scored on a scale of 0-2 based on the severity of the following clinical symptoms: grooming, posture, appearance of eyes and nose, breathing, dehydration, diarrhea, unprovoked behavior, and provoked behavior. Animals reaching endpoint criteria were humanely euthanized. Animal experiments were conducted in accordance with the Animal Ethics Review Committee of the University of Toronto.

FIG. 7 shows the results obtained. FIG. 7A shows a solid phase binding assay consisting of N.men cells fixed with paraformaldehyde (PFA) or lysed with SDS and were spotted onto nitrocellulose and probed with α-TbpB antibodies. ΔSLAM/tn5 refers to the original strain of SLAM deficient cells obtained through transposon insertion. ΔSLAM describes the knockout of SLAM in Neisseria meningitidis obtained by replacing the SLAM ORF with a kanamycin resistance cassette. FIG. 7B shows a Proteinase K digestion assay showing the degradation of TbpB, LbpB and fHbp only when Nm cells are SLAM deficient (ΔSLAM). Nm cells expressing individual SLPs alone and with SLAM were incubated with proteinase K and Western blots were used to detect levels of all three SLPs levels with and without protease digestion (−/+). Flow cytometry was used to confirm that ΔSLAM cells could not display TbpB (FIG. 7C) or fHbp (FIG. 7D) on the cell surface. Antibodies against TbpB and fHbp were used to bind surface exposed SLPs followed by incubation with a α-Rabbit antibody linked to phycoerythrin to provide fluorescence. The mean fluorescent intensity (MFI) of each sample was measured using the FL2 detector of a BD FACS Calibur. The signal obtained from wildtype cells was set to 100% for comparison with signals from knockout cells. Error bars represent the standard error of the mean (SEM) from three experiments. Shown in FIG. 7E are the results of mice infections with various strains. Mice were infected via intraperitoneal injection with 1×106 CFU of wildtype N. meningitidis strain B16B6, B16B6 with a knockout of TbpB (ΔtbpB), or B16B6 with a knockout of nmb0313 Δslam and monitored for survival and disease symptoms every 12 h starting 48 hr pre-infection to 48 h post-infection and additionally monitored at 3 hr post-infection. Statistical differences in survival were assessed by a Mantel-Cox log rank test (GraphPad Prism 5) (*p<0.05, n.s. not significant). These results show a marked reduction in post-infection mortality in mice infected with the knockout of nmb0313 Δslam strain.

US11872275B2. SLAM polynucleotides and polypeptides and uses thereof (2024-01-16). ENGINEERED ANTIGENS INC. [CA]. Inventors: Trevor F. Moraes [CA], Christine Chieh-Lin Lai [CA], Yogesh Hooda [CA], Andrew Judd [CA], Anthony B. Schryvers [CA], Scott D. Gray-Owen [CA].
Full text: Click here
Patent 2024
Agar Animals Antibodies Bacteremia Bacterial Infections Biological Assay Brain Cells Cultured Cells Dehydration Diarrhea Digestion Endopeptidase K Eye Flow Cytometry Fluorescence Genes Heart Homo sapiens Immunoglobulins Infection Injections, Intraperitoneal Jumping Genes Kanamycin Resistance Mice, Inbred C57BL Mus Neisseria Neisseria meningitidis Nitrocellulose Nose paraform Peptide Hydrolases Phycoerythrin prisma Rabbits Rivers Sepsis Strains Transferrin Virulence Western Blot
5
EV Protein Isolation and Western Blot
Total protein was isolated from EVs using Pierce RIPA lysis buffer with Halt protease and phosphatase inhibitor cocktail (Thermo Fisher, Waltham, MA), and protein concentration was determined using the Pierce BCA protein assay kit (Thermo Fisher). For Western blotting, samples were prepared in Laemmli buffer, boiled at 95 °C for 5 min, and 5 μg protein was loaded. Proteins were separated by SDS-PAGE using 4–15% TGX Gels (Criterion, Bio-Rad, Hercules, CA) by running at 200 V at room temperature. Proteins were transferred for 60 min at 100 V on ice onto a nitrocellulose membrane in 20% methanol Tris-glycine buffer. The Revert Total Protein Stain Kit (Li-Cor Biosciences, Lincoln, NE) or Ponceau S solution (Thermo Fisher) was used to stain total protein, and the membranes were imaged to verify transfer efficiency and loading. The membranes were subsequently blocked in 5% nonfat dry milk in Tris-buffered saline-Tween (TBS-T, 0.1% Tween-20) for 1 h at room temperature, then incubated overnight at 4 °C in primary antibody (anti-CD63, EXOAB-CD63A-1; System Biosciences, Palo Alto, CA and anti-Apolipoprotein A1 (ApoA1, 701239; Thermo Fisher) at a 1:1000 dilution in 5% nonfat dry milk in TBS-T. The membranes were then washed before incubation in goat anti-rabbit secondary antibodies (EXOAB-CD63A-1; System Biosciences) (1:10,000 dilution) for 1 h at room temperature. Blots were developed with enhanced chemiluminescence (Clarity Western ECL Substrate, Bio-Rad), imaged, and quantified with ImageJ (National Institutes of Health).
Ismaeel A., Van Pelt D.W., Hettinger Z.R., Fu X., Richards C.I., Butterfield T.A., Petrocelli J.J., Vechetti I.J., Confides A.L., Drummond M.J, & Dupont-Versteegden E.E. (2023). Extracellular vesicle distribution and localization in skeletal muscle at rest and following disuse atrophy. Skeletal Muscle, 13, 6.
Full text: Click here
Publication 2023
Anti-Antibodies APOA1 protein, human Biological Assay Buffers Chemiluminescence Gels Glycine Goat Immunoglobulins Laemmli buffer Methanol Milk, Cow's Nitrocellulose Peptide Hydrolases Phosphoric Monoester Hydrolases ponceau S Proteins Rabbits Radioimmunoprecipitation Assay Saline Solution SDS-PAGE Stains Technique, Dilution Tissue, Membrane Tween 20 Tweens

Top products related to «Nitrocellulose»

1
Nitrocellulose membrane by Bio-Rad
Sourced in United States, Germany, Italy, United Kingdom, Canada, France, China, Switzerland, Japan, Spain, Australia, Sweden, Portugal, Israel, Netherlands, Belgium
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.
2
Nitrocellulose membrane by Merck Group
Sourced in United States, Germany, China, United Kingdom, Ireland, France, Morocco, Italy, Australia, Sao Tome and Principe, Canada, Macao, Spain, India
Nitrocellulose membranes are a type of laboratory equipment used for various applications, such as protein blotting, DNA/RNA transfer, and immunodetection. They are made from purified cellulose that has been chemically treated to create a porous structure, allowing efficient binding and transfer of biomolecules. These membranes provide a reliable and versatile platform for a range of research and analytical procedures.
3
Nitrocellulose membrane by GE Healthcare
Sourced in United States, United Kingdom, Germany, Italy, Sweden, Switzerland, Australia, France, China, Canada, Japan, India
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.
4
Nitrocellulose membrane by Cytiva
Sourced in United Kingdom, United States, Germany, Sweden, Italy, France, Switzerland, Israel, Japan, China, Belgium, Canada
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.
5
Odyssey infrared imaging system by LI COR
Sourced in United States, Germany, United Kingdom, China, Italy, Niger, France, Netherlands, Austria, Australia, Liechtenstein
The Odyssey Infrared Imaging System is a versatile laboratory equipment designed for high-sensitivity detection and quantification of fluorescent and luminescent signals. The system utilizes infrared technology to capture and analyze various molecular targets, such as proteins, nucleic acids, and small molecules, in a range of sample types.
6
Protease inhibitor cocktail by Roche
Sourced in United States, Switzerland, Germany, China, United Kingdom, France, Canada, Japan, Italy, Australia, Austria, Sweden, Spain, Cameroon, India, Macao, Belgium, Israel
Protease inhibitor cocktail is a laboratory reagent used to inhibit the activity of proteases, which are enzymes that break down proteins. It is commonly used in protein extraction and purification procedures to prevent protein degradation.
7
Protease inhibitor cocktail by Merck Group
Sourced in United States, Germany, China, United Kingdom, Italy, Japan, Sao Tome and Principe, France, Canada, Macao, Switzerland, Spain, Australia, Israel, Hungary, Ireland, Denmark, Brazil, Poland, India, Mexico, Senegal, Netherlands, Singapore
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.
8
β actin by Merck Group
Sourced in United States, United Kingdom, Germany, China, Canada, Japan, Macao, Italy, Sao Tome and Principe, Israel, Spain, Denmark, France, Finland, Australia, Morocco, Ireland, Czechia, Sweden, Uruguay, Switzerland, Netherlands, Senegal
β-actin is a protein that is found in all eukaryotic cells and is involved in the structure and function of the cytoskeleton. It is a key component of the actin filaments that make up the cytoskeleton and plays a critical role in cell motility, cell division, and other cellular processes.
9
Nitrocellulose membrane by Thermo Fisher Scientific
Sourced in United States, United Kingdom, China, Germany, Switzerland, Sweden, Italy, Japan, Netherlands, Denmark
The Nitrocellulose membrane is a porous membrane made from cellulose treated with nitric acid. It is commonly used in various laboratory applications for the immobilization and detection of proteins, nucleic acids, and other biomolecules through techniques such as Western blotting, dot blotting, and nucleic acid hybridization.
10
Bca protein assay kit by Thermo Fisher Scientific
Sourced in United States, China, Germany, United Kingdom, Japan, Belgium, France, Switzerland, Italy, Canada, Australia, Sweden, Spain, Israel, Lithuania, Netherlands, Denmark, Finland, India, Singapore
The BCA Protein Assay Kit is a colorimetric detection and quantification method for total protein concentration. It utilizes bicinchoninic acid (BCA) for the colorimetric detection and quantification of total protein. The assay is based on the reduction of Cu2+ to Cu1+ by protein in an alkaline medium, with the chelation of BCA with the Cu1+ ion resulting in a purple-colored reaction product that exhibits a strong absorbance at 562 nm, which is proportional to the amount of protein present in the sample.

More about "Nitrocellulose"

Nitrocellulose, a highly flammable compound derived from the nitration of cellulose, has a wide range of applications, including in lacquers, explosives, and as a binder in paints and photographic films.
Researchers working with nitrocellulose must engage in careful planning and execution to ensure reproducibility and accuracy in their studies.
PubCompare.ai, the leading AI-driven platform, can assist researchers in optimizing their nitrocellulose research by easily locating the best protocols from literature, pre-prints, and patents using advanced AI-powered comparisons.
This can help researchers discover the most reliable and effective nitrocellulose-related products and methodologies, elevating their research outcomes.
Nitrocellulose membranes, commonly used in Western blotting and other analytical techniques, are an important component in nitrocellulose-based research.
The Odyssey Infrared Imaging System can be utilized to visualize and quantify the proteins separated on nitrocellulose membranes, while protease inhibitor cocktails help preserve the integrity of the proteins during the process.
Additionally, the use of β-actin as a loading control is a common practice in nitrocellulose-based research, as it helps to normalize the protein levels across samples.
The BCA protein assay kit can be employed to accurately quantify the protein concentrations prior to loading onto the nitrocellulose membrane.
Experience the power of PubCompare.ai today and take your nitrocellulose research to new hights.