The liver tissues were isolated and pestled, and were lysed in radioimmunoprecipitation assay buffer. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis was performed to separate equal amounts of cell lysates, and were transferred to a polyvinylidene difluoride membrane (Millipore, Burlington, MA). After incubation with the primary antibodies overnight at 4°C, and corresponding secondary antibodies for 2 hours at room temperature. Finaly, dropped electrochemiluminescence liquid for 1 minute, signals were detected using a chemiluminescence instrument (Imagequant LAS500, Cytiva, USA). Primary antibodies were as follows: CMV-IE1 and IE2 (1:500, cat. ab53495; Abcam, Cambridge, UK), P27 (1:1000, cat. A16722; ABclonal, Wuhan, China), P21 (1:1000, cat. bs-55160R; Bioss, Beijing, China), CEBPα (1:1000, cat. A0904; ABclonal, Wuhan, China), HNF4α (1:1000, cat. A13998; ABclonal, Wuhan, China), and β-actin (1:2000; Santa Cruz, CA). The secondary antibodies used were horseradish peroxidase–conjugated sheep anti-mouse IgG (1:500; Absin, Shanghai, China) and anti-rabbit (1:10,000; Absin, Shanghai, China).
Hnf4α
Manufactured by ABclonal
Sourced in China
HNF4α is a transcription factor that plays a crucial role in the regulation of gene expression. It is involved in the control of various cellular processes, including cell differentiation, metabolism, and development. HNF4α is an important regulator of hepatocyte-specific gene expression and is essential for liver function and development.
Automatically generated - may contain errors
Lab products found in correlation
Polyvinylidene difluoride membrane, by Merck Group (1 mentions)
β actin, by Santa Cruz Biotechnology (1 mentions)
Cmv ie1 and ie2, by Abcam (1 mentions)
Horseradish peroxidase conjugated sheep anti mouse igg, by Absin (1 mentions)
C ebpα, by Bioss Antibodies (1 mentions)
Imagequant las 500, by Cytiva (1 mentions)
P akt ser473, by Cell Signaling Technology (1 mentions)
Goat anti rabbit hrp conjugated secondary antibody, by Bio-Rad (1 mentions)
P akt thr308, by Cell Signaling Technology (1 mentions)
Gsk3α β, by Cell Signaling Technology (1 mentions)
P gsk3β ser9, by Cell Signaling Technology (1 mentions)
P erk thr202 tyr204, by Cell Signaling Technology (1 mentions)
P gs ser641, by Cell Signaling Technology (1 mentions)
P irs 1 ser307, by Cell Signaling Technology (1 mentions)
P irs 1ser612, by Cell Signaling Technology (1 mentions)
P foxo1 ser256, by Cell Signaling Technology (1 mentions)
Foxo3a, by Cell Signaling Technology (1 mentions)
Foxo1, by Cell Signaling Technology (1 mentions)
β actin, by ABclonal (1 mentions)
Pgc 1α, by ABclonal (1 mentions)
2 protocols using hnf4α
1
Liver Protein Expression Analysis
2
Western Blot Analysis of Insulin Signaling
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The tissues were isolated from mice in the fed condition and then immediately frozen in liquid nitrogen. Tissue homogenates were prepared, and 30–50 μg of protein was resolved on 4%–12% SDS-PAGE gels and subjected to Western blotting (12 (link)). Immunoblots were performed using antibodies against the following proteins: AKT (catalog 4685), P-AKTThr308 (catalog 13038), P-AKTSer473 (catalog 4060), FoxO1 (catalog 2880), FoxO3a (catalog D19A7), P-FoxO1Ser256 (catalog 9461), P-FoxO1Thr24/FoxO3aThr32/FoxO4Thr28 (catalog 2599), FoxO1 (catalog 2880), GSK3α/β (catalog 5676), P-GSK3βSer9 (catalog 5558), GS (catalog 3893), P-GSSer641 (catalog 94905), P-IRS-1Ser307 (catalog 2381), P-IRS-1Ser612 (catalog 3203), P-TSC2Ser939 (catalog 3615), TSC2 (catalog 3612), p-mTORSer2449 (catalog 5536), mTOR (catalog 2983), and P-ERKThr202/Tyr204 (catalog 9101) were purchased from Cell Signaling Technology. Antibodies against HNF-4α (catalog A2085), PGC-1α (catalog A11971), GK (catalog Ab293), G6Pase (catalog A16234), GKCR (catalog A5678), and β-actin (catalog AC026) were purchased from ABclonal, and ERK (catalog 514302) was purchased from Santa Cruz Biotechnology. Goat anti-mouse and goat anti–rabbit HRP–conjugated secondary antibodies (1:3000; Bio-Rad, 1662408EDU) were used for described experiments.
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