Nitrocellulose membrane

Manufactured by Genesee Scientific

Sourced in United States

Nitrocellulose membranes are porous sheets used for the immobilization and detection of biomolecules, such as proteins and nucleic acids, in various laboratory applications. They provide a stable and inert surface for the binding and analysis of these biomolecules.

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Lab products found in correlation

Goat anti human fc specific antibody, by Merck Group (2 mentions) Goat anti human igg h l antibodies, by Jackson ImmunoResearch (2 mentions) C digit blot scanner, by LI COR (2 mentions) Human serum, by Merck Group (2 mentions) Supersignal west pico chemiluminescent substrate, by Thermo Fisher Scientific (2 mentions) Gradient polyacrylamide gel, by Bio-Rad (1 mentions) Anti v5 antibody, by Rockland Immunochemicals (1 mentions) Anti alpha tubulin, by Developmental Studies Hybridoma Bank (1 mentions)

4 protocols using nitrocellulose membrane

Antibody Depletion and Detection in Human Serum

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Human serum (Sigma-Aldrich, St. Louis, MO, USA) was depleted of
endogenous IgG by passage through a protein G-Sepharose column. Antibodies and
ADCs were incubated at a concentration of 40 μg/ml in IgG-depleted serum
for 0, 3, or 5 days at 37 °C followed by immunoprecipitation using
agarose beads coupled to goat anti-human Fc-specific antibody (Sigma-Aldrich).
Immunoprecipitated antibodies and ADCs were run on SDS-polyacrylamide gels and
transferred onto nitrocellulose membranes (0.45 μm pore size; Genesee
Scientific, San Diego, CA, USA). Immunoblotting was carried out using standard
methods with goat anti-human IgG (H + L) antibodies conjugated with HRP (Jackson
Immunoresearch, West Grove, PA, USA). HRP was detected using SuperSignal West
Pico Chemiluminescent Substrate (Thermo Fisher Scientific) followed by scanning
with a C-DiGit blot scanner (LI-COR Biosciences, Lincoln, NE, USA).

Kang J.C., Sun W., Khare P., Karimi M., Wang X., Shen Y., Ober R.J, & Ward E.S. (2019). Engineering a HER2-specific antibody-drug conjugate to increase lysosomal delivery and therapeutic efficacy. Nature biotechnology, 37(5), 523-526.

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Antibody Depletion and Detection in Human Serum

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Immunoblotting of aPKC and β-tubulin

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ASZ001 cells were seeded to confluency, serum-starved and treated with DMSO or Everolimus (10 nM) for 24 h. Cells were collected and lysed in SDS sample preparation buffer (100 mM Tris-HCl, pH 6.8; 1 M DTT, 4% SDS, 20% glycerol and 0.2% bromophenol blue). Samples were loaded onto a 4–20% gradient polyacrylamide gel (Bio-Rad) and transferred onto nitrocellulose membrane (Genesee Scientific). Membranes were immunoblotted with antibodies against aPKC (Santa Cruz Biotech, 1:1000) and β-tubulin (DSHB, 1:2000) in 1× TBST, incubated with secondary antibodies donkey anti-mouse Alexa 680 or donkey anti-rabbit Alexa 790 (Jackson, 10 000), and then imaged using the LI-COR Odyssey System.

Chow R.Y., Levee T.M., Kaur G., Cedeno D.P., Doan L.T, & Atwood S.X. (2020). MTOR promotes basal cell carcinoma growth through atypical PKC. Experimental dermatology, 30(3), 358-366.

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Investigating MRAP Interactions with MC2R Mutants

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CHO cells were separately transfected with either wild-type xtMC2R or xtMC2R mutant I158/A, F161/A (TM4), V172/A (TM5) or M166/A (EC2) and each can was co-expressed with cMRAP for 5 days in advance of cell lysis. Cells were homogenized in RIPA buffer and sonicated to collect total protein. Sample protein levels were then measured by nano-spectrophotometry and analyzed by poly-acrylamide gel electrophoresis. A 4% to 20% Mini-PROTEAN® TGX™ Precast Protein Acrylamide Gels (Biorad, Hercules, CA, USA), and then samples were transferred to a nitrocellulose membrane (Genesee Scientific; San Diego, CA, USA). Membrane was blotted with anti-V5 antibody (Rockland Immunochemicals, Limerick, PA, USA) at a final concentration of 1:500, and anti-alpha-tubulin from Developmental Studies Hybridoma Bank (University of Iowa) at a final concentration of 1:500. Membrane was scanned using a Protein Simple FluorChem system (San Jose, CA, USA) and analyzed using Fiji; an open-source platform for biological-image analysis. The Western analysis was done in triplicate.

Davis P.E., Wilkinson E.C, & Dores R.M. (2019). Identifying Common Features in the Activation of Melanocortin-2 Receptors: Studies on the Xenopus tropicalis Melanocortin-2 Receptor. International Journal of Molecular Sciences, 20(17), 4166.

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