Hrp conjugated gapdh

Manufactured by Cell Signaling Technology

HRP-conjugated GAPDH is a horseradish peroxidase (HRP)-labeled antibody that specifically binds to the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein. GAPDH is a ubiquitous enzyme involved in glycolysis and is often used as a loading control in Western blot analysis.

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Lab products found in correlation

P akt ser473, by Cell Signaling Technology (1 mentions) P erk, by Cell Signaling Technology (1 mentions) P akt thr308, by Cell Signaling Technology (1 mentions) Anti braf, by Abcam (1 mentions) Anti erk1 2, by Promega (1 mentions) P mek, by Cell Signaling Technology (1 mentions) Image lab software, by Bio-Rad (1 mentions) Pvdf membrane, by Merck Group (1 mentions) Ripa buffer, by Cell Signaling Technology (1 mentions) Bolt 4 12 bis tris gradient gels, by Thermo Fisher Scientific (1 mentions) Protease inhibitor cocktail, by Roche (1 mentions) Chemidoc mp imaging system, by Bio-Rad (1 mentions) Anti ha tag, by Cell Signaling Technology (1 mentions)

Spelling variants of this product

GAPDH (5572 citations) GAPDH-HRP (14 citations) HRP conjugated anti-GAPDH (5 citations) Rabbit anti-GAPDH conjugated to horseradish peroxidase (HRP) (2 citations)

2 protocols using hrp conjugated gapdh

Immunoblotting Analysis of MAPK and PI3K/mTOR Pathways

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Protein concentrations of cell samples were determined by using Pierce 660nm Protein Assay and run on NuPAGE precast gels (4%-12% Bis-Tris or Tris-acetate). Immunoblots were incubated with the indicated primary antibodies per manufactures’ recommendations and as described previously [4 (link)]. For MAPK and PI3K/mTOR pathway analysis, pMEK (#9154), MEK (#4694), pERK (#4370), ERK (#4695), pAKT Ser473 (#4060), pAKT Thr308 (#4056), AKT (#2920), pS6 (#2215) and S6 (#2317) antibodies from Cell Signaling were used. HRP-conjugated Beta-Actin, and HRP-conjugated GAPDH were also obtained from Cell Signaling Biotechnology, Inc.; anti-ERK 1/2 was obtained from Promega; anti-BRAF was obtained from Abcam.

Jain P., Silva A., Han H.J., Lang S.S., Zhu Y., Boucher K., Smith T.E., Vakil A., Diviney P., Choudhari N., Raman P., Busch C.M., Delaney T., Yang X., Olow A.K., Mueller S., Haas-Kogan D., Fox E., Storm P.B., Resnick A.C, & Waanders A.J. (2017). Overcoming resistance to single-agent therapy for oncogenic BRAF gene fusions via combinatorial targeting of MAPK and PI3K/mTOR signaling pathways. Oncotarget, 8(49), 84697-84713.

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Western Blot Protein Detection

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Cells were lysed in 1xRIPA buffer (Cell Signaling Technology) supplemented with protease inhibitor cocktail (Roche). Equal volumes cell lysate were run on BOLT 4–12% Bis-Tris gradient gels (Invitrogen) and transferred to PVDF membranes (Millipore). Non-specific antigen binding was blocked with TBS-T (50mM Tris, 150mM NaCl and 0.05% Tween-20) with 5% BLOT-QuickBlocker Reagent (Millipore) for 1 hour. Membranes were incubated with primary antibodies (anti-HA-tag (Cell Signaling Technology C29F4) or HRP-conjugated GAPDH (Cell Signaling Technology 14C10)) for 1 hour in 1% BLOT-QuickBlocker. Membranes were washed for 3 10 minute washes and anti-HA-tag membranes were further incubated with anti-rabbit antibody (Cell Signaling Technology 7074) for 1h followed by 6 10 minute washes in TBS-T. Proteins were visualized with West Pico Chemiluminescent Substrate (Life Technology) and imaged using the ChemiDoc MP Imaging System (Bio-Rad) and processed with ImageLab software (Bio-Rad).

Zetsche B., Gootenberg J.S., Abudayyeh O.O., Slaymaker I.M., Makarova K.S., Essletzbichler P., Volz S., Joung J., van der Oost J., Regev A., Koonin E.V, & Zhang F. (2015). Cpf1 is a single RNA-guided endonuclease of a Class 2 CRISPR-Cas system. Cell, 163(3), 759-771.

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