Protein isolation was performed through cell lysis buffer (R0278-50ML, Sigma-Aldrich Co., USA) with their concentrations quantified by Bradford reagent (B6916-500ML, Sigma-Aldrich Co., USA). After boiling, 30 μg of proteins were separated by polyacrylamide gel electrophoresis and transferred to a Hybond ECL nitrocellulose membrane (Amersham Biosciences, Arlington Heights, IL, USA). Membranes were then blocked with 5% low-fat dried milk in phosphate buffered saline containing 0.1% Tween-20 and subsequently incubated for 1 h at room temperature with 1:1,000 dilutions of the primary ZNF703 antibody. Mouse monoclonal anti-GAPDH (KC5G4, KangChen Biotech, Shanghai, China) was used as a loading control. Membranes were then exposed to ECL western blotting substrate (32209, Thermo Fisher Scientific Inc. Waltham, USA) after incubating for 1 h with 1:1,000 dilution of the second antibody in darkness.
Anti gapdh kc5g4
Manufactured by Kangchen Biotech
Sourced in China
Anti-GAPDH (KC5G4) is a monoclonal antibody that binds to the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein. GAPDH is a widely expressed and highly conserved enzyme involved in the glycolytic pathway. This antibody can be used to detect and quantify GAPDH levels in various research applications.
Automatically generated - may contain errors
Lab products found in correlation
R0278 50ml, by Merck Group (1 mentions)
Ecl western blotting substrate, by Thermo Fisher Scientific (1 mentions)
Hybond ecl nitrocellulose membrane, by Cytiva (1 mentions)
Bradford reagent, by Merck Group (1 mentions)
Quantity one software version 4, by Bio-Rad (1 mentions)
Micro bca assay kit, by Thermo Fisher Scientific (1 mentions)
M per mammalian protein extraction reagent, by Thermo Fisher Scientific (1 mentions)
2 protocols using anti gapdh kc5g4
1
Quantitative Protein Analysis by Western Blot
2
Protein Extraction from Heat-Stressed H9c2 Cells
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Heat-stressed H9c2 cell protein was extracted using M-PER Mammalian Protein Extraction Reagent (78501; Thermo Scientific, Waltham, MA, USA) according to the manufacturer protocol. Heart tissue samples were homogenized in 1 mL protein extraction reagent on ice using an Ultra-Turrax homogenizer after being completely washed in ice-cold physiological saline; the homogenates were then centrifuged at 12,000 g for 20 min at 4°C to remove cellular debris. The supernatant was collected and stored at -20°C for protein quantification. Protein concentrations were measured using a Micro BCA assay kit (23235; Thermo Scientific). The bands on the developed film were quantified using the Quantity One software version 4.6.2 (Bio-Rad, Hercules, CA, USA). All blots were probed with anti-GAPDH (KC-5G4; KangChen Bio-tech Inc., Shanghai, China) antibody as an internal control.
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