The cells were lysed in RIPA buffer (Solarbio, R0010). The protein concentration was determined using the BCA Protein Assay Kit (Meilunbio, MA0082). Equivalent amounts of protein were separated through SDS-polyacrylamide gels and electroblotted onto PVDF membranes (Bio-Rad, 1620177). After blocking using a 5% BSA Blocking Buffer (Solarbio, SW3015), the membranes were incubated with the primary antibody at 4 °C overnight. The antibodies used were AMPKα Antibody (Cell Signaling Technology, 2532, Danvers, MA, USA), Phospho-AMPKα Rabbit mAb (Cell Signaling Technology, 2535), mTOR Antibody (Cell Signaling Technology, 2972), Phospho-mTOR Antibody (Cell Signaling Technology, 2971), LC3B (E5Q2K) Mouse mAb (Cell Signaling Technology, 83506), SQSTM1/p62 Antibody (Cell Signaling Technology, 5114), and β-Actin Antibody (Cell Signaling Technology, 4967). Finally, the relevant protein was visualized through staining with an appropriate HRP-conjugated secondary antibody for 1 h and then enhanced with chemiluminescence. The expression of the protein was quantified using ImageJ software.
Phospho ampkα rabbit mab
Manufactured by Cell Signaling Technology
Sourced in United States
Phospho-AMPKα Rabbit mAb is a laboratory reagent used for the detection and analysis of phosphorylated AMP-activated protein kinase alpha (AMPKα) in various experimental systems.
Automatically generated - may contain errors
Lab products found in correlation
Sqstm1 p62 antibody, by Cell Signaling Technology (1 mentions)
Mtor antibody, by Cell Signaling Technology (1 mentions)
Ampkα antibody, by Cell Signaling Technology (1 mentions)
Bca protein assay kit, by Meilun (1 mentions)
Pvdf membrane, by Bio-Rad (1 mentions)
Bsa blocking buffer, by Solarbio (1 mentions)
Ripa buffer, by Solarbio (1 mentions)
β actin antibody, by Cell Signaling Technology (1 mentions)
Lc3b e5q2k mouse mab, by Cell Signaling Technology (1 mentions)
Phospho mtor antibody, by Cell Signaling Technology (1 mentions)
Fas mouse mab, by Santa Cruz Biotechnology (1 mentions)
Ecl western blotting detection reagent, by Cytiva (1 mentions)
Gel pro analyzer 4, by Media Cybernetics (1 mentions)
2 protocols using phospho ampkα rabbit mab
1
Western Blot Analysis of AMPK and mTOR
2
Western Blotting Analysis of AMPK Pathway
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Liver and adipose tissue protein extraction and Western blotting were performed as described before (21 (link)). The primary antibodies were comprised of AMPKα rabbit mAb (cat. No. 5831, Cell Signaling, MA, USA), phospho-AMPKα rabbit mAb (cat. No. 50081, Cell Signaling, MA, USA), ACC rabbit mAb (cat. No. 3676, Cell Signaling, MA, USA, phospho-ACC rabbit mAb (cat. No. 11818, Cell Signaling, MA, USA), CPT1A rabbit pAb (cat. No. 15184-1-ap, Proteintech, USA), SREBP-1 mouse mAb (cat. No. sc-13551, Santa Cruz Biotechnology), peroxisome proliferation-activated receptor (PPAR)-α mouse mAb (cat. No. sc-398394, Santa Cruz Biotechnology) and FAS mouse mAb (cat. No. sc-55580, Santa Cruz Biotechnology). Primary antibody binding was detected using anti-rabbit or anti-mouse secondary antibodies conjugated with horseradish peroxidase (cat. No. 111-035-003, Jackson; cat. No. 115-035-003, Jackson). The expression of antibody linked protein was determined by ECL™ Western Blotting Detection Reagents (Amersham Pharmacia Biotech Inc., NJ, USA). The optical density of the bands was quantified by Gel-Pro Analyzer 4 (Media Cybernetics Corporation, USA).
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