Tissue samples were homogenized and protein concentrations determined according to the method of Bradford protein assay. The proteins of equal loading were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and transferred to a polyvinylidene difluoride membrane. Membranes were incubated with rabbit monoclonal anti‐S100A6 antibody (Novus Biologicals, Littleton, CO; catalog #NBP1‐95284) or rabbit polyclonal anti–green fluorescent protein (GFP) antibody (Abcam, Cambridge, UK; catalog #ab6556) and anti‐rabbit IgG horseradish peroxidase–conjugated (Promega, Madison, WI; catalog #W401B). Blots of proteins were probed with rabbit polyclonal anti‐GAPDH primary antibody (Santa Cruz Biotechnologies, Dallas, TX; catalog #sc‐25778) for internal reference. Secondary antibodies were detected using Immobilon Western chemiluminescence horseradish peroxidase substrate (Millipore, Billerica, MA; catalog #WBKLS0050). Band intensity was quantified by scanning densitometry using a VersaDoc Imaging System (Bio‐Rad, Hercules, CA).
Anti gapdh primary antibody
Manufactured by Santa Cruz Biotechnology
Sourced in United States
The Anti-GAPDH primary antibody is a tool used in scientific research to detect and study the GAPDH protein, which is involved in various cellular processes. This antibody specifically binds to and identifies the GAPDH protein, allowing researchers to analyze its presence and behavior in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Versadoc imaging system, by Bio-Rad (1 mentions)
Immobilon western chemiluminescence horseradish peroxidase substrate, by Merck Group (1 mentions)
Trans blot turbo transfer system, by Bio-Rad (1 mentions)
Sc 47724, by Santa Cruz Biotechnology (1 mentions)
Tunel kit, by Roche (1 mentions)
Appropriate secondary antibody, by Santa Cruz Biotechnology (1 mentions)
Anti bax, by Abcam (1 mentions)
Anti caspase 3, by Abcam (1 mentions)
Anti bcl 2, by Abcam (1 mentions)
Anti nphs2, by Abcam (1 mentions)
Anti tgf β1, by Abcam (1 mentions)
Bun kit, by Nanjing Jiancheng (1 mentions)
Anti gli1 antibody, by Abcam (1 mentions)
4 protocols using anti gapdh primary antibody
1
Western Blot Analysis of S100A6 and GFP Proteins
2
Immunoblotting Analysis of DARTS Protein
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Immunoblotting analyses were carried out by submitting 20 μg of the DARTS protein mixtures to 1D-SDS-PAGE, then transferring the gels onto nitrocellulose membranes through the Trans-Blot Turbo Transfer System (Bio-Rad, Hercules, CA, USA) for 15 min at 2.5 A (up to 25 V). The membranes were blocked for 1 h at room temperature in a 5% w/vol non-fat dried milk solution and then incubated overnight at 4 °C with the following primary antibodies: anti-USP-7 (SC-137008, Santa Cruz Biotechnology, Dallas, TX, USA) 1:250 vol/vol, anti-PES (SC-166300, Santa Cruz Biotechnology, Dallas, TX, USA) 1:500 vol/vol, anti-NXF1 (SC-32319, Santa Cruz Biotechnology, Dallas, TX, USA) 1:500 vol/vol, then mouse peroxidase-conjugated secondary antibody was added (1:500 vol/vol Thermo-Scientific, Bremen, Germany). Membranes were also hybridized with an anti-GAPDH primary antibody (SC-47724, Santa Cruz Biotechnology, Dallas, TX, USA) 1:2000 vol/vol) for loading normalization purposes.
3
Renal Protection by Microbubble-Mediated Drug Delivery
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Egg Yolk Lecithin (PC, PC-98T) and cholesterol (98% of purity) were purchased from Shanghai Advanced Vehicle Technology Pharmaceutical L.T.D. Co (AVT). Anti-Bax, anti-Bcl-2, anti-NPHS2, anti-caspase-3 and anti-TGFβ1 antibodies were obtained from Abcam (Abcam, Cambridge, MA). Anti-GAPDH primary antibody, and appropriate secondary antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). CRE, TG and BUN kit were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing Jiancheng Bioengineering Institute, Nanjing, China). TUNEL kit was obtained from Roche (Roche, Shanghai, China). All other reagents were obtained from Sigma-Aldrich (Sigma-Aldrich, St. Louis, MO). Simulated body fluid (SBF) were purchased from Beijing leagene Biotechnology Company (Beijing leagene Biotechnology Company, Beijing, China). MB was prepared by the sonication-lyophilization method as reported in our recent study [46 (link)].
4
Characterization of GLI1 Protein Expression
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AtT-20 cells were harvested after 48 h and proteins were isolated using radioimmunoprecipitation assay (RIPA) lysis buffer containing protease and phosphatase inhibitors (cOmplete, Cat. No. 04693124001 and PhosSTOP, Cat. No. 04906845001, both from Roche). Precast tris glycine gels were loaded with 20 mg of proteins and transferred to a PVDF membrane. The membrane was blocked with 1% BSA, and anti-GLI1 antibody (rabbit polyclonal, Abcam, Cat. No. ab151796; 1:700) was incubated over night at 4 8C. Secondary antibody (rabbit polyclonal, Dako, Glostrup, Denmark, Cat. No. P0448; 1:4000) was incubated for 1 h at RT. For GAPDH, anti-GAPDH primary antibody (rabbit polyclonal, Santa Cruz; Cat. No. sc-25778; 1:3000) and the above secondary antibody were used.
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