Enzo Biochem Inc.
Anti-GAPDH antibody
ENZ-ABS276
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The expression levels of GAPDH were used as internal loading control. Western blot data using this antibody has been published in a paper.
Western Blot
Cell lysates
1/1000
5% milk
1/2000
Room temperature 1 hour
ECL
Lysates were made by adding lysis buffer [1% SDS, 10 mmol/L Tris-Cl (pH 7.6), 20 μg/μL aprotinin, 20 μg/μL leupeptin, and 1 mmol/L 4-(2-aminoethyl)benzenosulfonyl fluoride]. The supernatants were obtained after centrifugation at 12000 g for 15 minutes at 4. The protein concentrations were determined using the Bicinchoninic Acid Protein Assay kit (Pierce, Rockford, IL). Fiften micrograms of protein were separated on SDS-PAGE gels and transferred to polyvinylidene difluoride membranes. After blocking, the membranes were incubated with the appropriately diluted primary antibody at 4°C overnight. After washing with TBST, the membranes were incubated with appropriately diluted HRP-conjugated secondary antibodies at room temperature for 1 hour. Proteins were detected with the enhanced chemiluminescence kit (Amersham Pharmacia Biotechnology, Inc., Piscataway, NJ).
https://doi.org/10.1186/s12885-018-4965-6
Figure 1A
Worked well
N/A
Worked as expected