Adipose tissue samples were homogenized in RIPA lysis buffer containing protease inhibitors. Protein concentrations were determined by the BCA method. Proteins (110 μg) were separated on 10% SDS-PAGE gels and transferred to a PVDF membrane (Millipore, USA).The membranes were blocked in 5% (w/v) non-fat milk for 1 h and then incubated with rabbit anti-ATGL (Cell Signaling Beverly, MA, USA,1:1000 dilution), rabbit anti-GAPDH (CW Biotech, Beijing, China, 1:3000 dilution) or mouse anti-β-actin (Proeintech, Chicago, IL, USA, 1:2000 dilution) primary antibodies overnight at 4°C. Subsequently, the membranes were incubated with peroxidase-conjugated anti-rabbit or anti-mouse secondary antibody for 1 h at room temperature. After washing with TBST, the immune complexes were detected with the Alpha Q Chemiluminescence System and exposed to film. The relative intensity of the target protein to GAPDH or to β-actin in the same sample was analyzed with Alpha Q software.
Rabbit anti gapdh
Manufactured by CWBIO
Sourced in United States
Rabbit anti-GAPDH is a primary antibody that specifically recognizes 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 expression in various sample types.
Automatically generated - may contain errors
2 protocols using rabbit anti gapdh
1
Western Blot Analysis of Adipose Tissue
2
Penile Tissue Protein Analysis
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The cellular protein samples were prepared by homogenization of penile tissue in a lysis buffer containing 1% IGEPAL CA-630, 0.5% sodium deoxycholate, 0.1% sodium docecyl sulfate, aprotinin (10 mg/mL), leupeptin (10 mg/mL), and phosphate buffered saline. The cellular lysates from penis containing 20 mg of protein were electrophoresed in sodium dodecyl sulfate–polyacrylamide gel electrophoresis and then transferred to a polyvinylidene fluoride membrane (Millipore Corp.). Primary antibodies were rabbit anti-⍺-SMA (1:1000; Abcam), rabbit anti- nNOS (1:400; Abcam) and rabbit anti-GAPDH (1:2000; cwBio, Lot:01225/50,404). After the hybridization of secondary antibodies, the resulting images were analyzed with ChemiImager 4000 (Alpha Innotech Corporation, San Leandro, CA, USA).
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