Cell lysates were prepared in lysis buffer containing 0.5% Nonidet P-40 as described.12 (link) The total protein were quantified and then boiled in 4 × SDS sample buffer followed by separation on SDS polyacrylamide gels. Immunoblotting was performed with the antibodies, as follows: anti-Flag, anti-β-actin, anti-mouse secondary and anti-rabbit secondary antibodies (Sigma); anti-Myc, anti-phospho-AMPK and anti-AMPK (Cell signaling Technology); anti-Ubiquitin, anti-α-Tubulin and anti-EGFP (Santa Cruz Inc.); and anti-HIF-1α, anti-Itch (BD Transduction Laboratories). Ubiquitin-immunoprecipitation assay was performed as described.12 (link) Size of bands are indicated in the blots.
Anti phospho ampk and anti ampk
Manufactured by Cell Signaling Technology
Anti-phospho-AMPK and anti-AMPK are antibodies produced by Cell Signaling Technology. These antibodies are used to detect the phosphorylated and total forms of AMP-activated protein kinase (AMPK), respectively. AMPK is a key cellular energy sensor that plays a crucial role in regulating cellular metabolism.
Automatically generated - may contain errors
Lab products found in correlation
Anti myc, by Cell Signaling Technology (1 mentions)
Anti flag, by Merck Group (1 mentions)
Anti β actin, by Merck Group (1 mentions)
Anti β actin antibody, by Santa Cruz Biotechnology (1 mentions)
Anti phospho akt and anti akt, by Cell Signaling Technology (1 mentions)
Anti srebp 1c, by Santa Cruz Biotechnology (1 mentions)
2 protocols using anti phospho ampk and anti ampk
1
Ubiquitin-mediated Signaling Pathway Analysis
2
Transcriptional Regulation of Lipid Metabolism
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The RT-qPCR analyses of SREBP-1c, FAS, ACC, PPARα, ACOX and CPT1 were performed as described previously (Park et al. 2016) (link). The quantity of each target mRNA was quantified relative to 36B4 mRNA. The ChIP assays were performed using HepG2 cells as described previously using the anti-SREBP-1c (Santa Cruz) or anti-PPARα antibodies (Abcam). Sequences of primer pairs used for real-time PCR are listed in Table 2 (Park et al. 2016) (link). The immunoblot analyses were performed as described previously using the anti-SREBP-1c (Santa Cruz), anti-phospho-PPARα and anti-PPARα (Abcam), anti-phospho-AMPK and anti-AMPK (Cell Signaling), anti-phospho-ACC and anti-ACC (Cell Signaling), anti-phospho-Akt and anti-Akt (Cell Signaling) and anti-β-actin antibodies (Santa Cruz). The level of each protein was quantified using the ImageJ program and normalized to the level of β-actin.
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