The isolated protein samples were subjected to 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto nitrocellulose filter membranes (Millipore, Bedford, MA, USA). Afterward, the membranes were then incubated with primary antibodies for a whole night, including Bcl-2, PCNA, TGFB2, Smad2, p-Smad2, and glyceraldehydes 3-phosphate dehydrogenase (1:1,000; Cell Signaling Technology, Beverly, MA, USA), followed by probing with corresponding horseradish peroxidase (HRP)-conjugated secondary antibodies (1:3,000, Abcam) for 1 h at 37°C. Protein bands were detected by enhanced chemiluminescence (ECL) plus western blotting detection reagents (GE Healthcare Life Sciences, Piscataway, NJ, USA) and analyzed by Quantity One analysis software (Bio-Rad, San Francisco, CA, USA).
Glyceraldehydes 3 phosphate dehydrogenase
Manufactured by Cell Signaling Technology
Sourced in United States
Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is an enzyme involved in the glycolytic pathway, catalyzing the reversible oxidation of glyceraldehyde 3-phosphate to 1,3-bisphosphoglycerate. It plays a crucial role in energy production within cells.
Automatically generated - may contain errors
Lab products found in correlation
Quantity one analysis software, by Bio-Rad (1 mentions)
Western blotting detection reagent, by GE Healthcare (1 mentions)
Horseradish peroxidase hrp conjugated secondary antibody, by Abcam (1 mentions)
Proliferating cell nuclear antigen (pcna), by Cell Signaling Technology (1 mentions)
Bcl 2, by Cell Signaling Technology (1 mentions)
Nitrocellulose filter membrane, by Merck Group (1 mentions)
Tgfb2, by Cell Signaling Technology (1 mentions)
Smad2, by Cell Signaling Technology (1 mentions)
P smad2, by Cell Signaling Technology (1 mentions)
Phospho akt ser473, by Cell Signaling Technology (1 mentions)
2 protocols using glyceraldehydes 3 phosphate dehydrogenase
1
Western Blot Analysis of Cellular Proteins
2
Insulin Stimulation Protein Analysis
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Insulin stimulation in vivo was performed with 5 units per kg insulin injected intraperitoneally. Tissues were harvested 10 min after injection. Protein lysates were separated by SDS–PAGE and immunoblotted with antibodies for FAK (sc-557), Fas (sc-1023), Fas ligand (sc-834-G), cyclin-dependent kinase 55 (sc-173), cleaved caspase 3 (a-277) (Santa Cruz Biotechnology, Inc.), Akt (4691), phospho-Akt (Ser473) (9271), phospho-ERK1/2 (9101), perilipin (3470), phospho-p53 (12571), p53 (2524S), glyceraldehydes-3-phosphate dehydrogenase (2118) and phospho-FAK (Tyr397) (3283) (Cell Signaling). Primary antibodies were diluted 1:1,000 and secondary antibodies diluted 1:3,000. Band intensities were quantified using ImageJ software61 (link). Uncropped images of Western blots are available in Supplementary Fig. 12 .
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