Total protein was extracted from liver tissues or HepG2 cells. Samples were separated using sodium dodecyl sulphate‐polyacrylamide gel electrophoresis and electroblotted onto polyvinylidene difluoride membranes. The membranes were first blocked with non‐fat milk for 2 h, and incubated with the following primary antibodies: anti‐PPARγ, anti‐PPARα, anti‐CD36(ABclonal Technology, Boston, MA, USA), anti‐GAPDH and anti‐α‐tubulin (Beijing Ray antibody, Beijing, China), followed by incubation with a horseradish peroxidase‐conjugated anti‐rabbit or mice secondary antibody (Beijing Ray antibody). Objective proteins were detected using ECL Plus (Pierce, NJ, USA).
Anti cd36
Manufactured by ABclonal
Sourced in United States
Anti-CD36 is a lab equipment product used for the detection and analysis of CD36, a cell surface receptor involved in various biological processes. It provides researchers with a reliable tool for identifying and quantifying CD36 expression in different cell types and samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti gapdh, by ABclonal (2 mentions)
Anti pparα, by ABclonal (1 mentions)
Anti pparγ, by ABclonal (1 mentions)
Anti ikbα, by Proteintech (1 mentions)
Anti β actin, by ABclonal (1 mentions)
Anti fto, by ABclonal (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
Anti pparγ, by Cell Signaling Technology (1 mentions)
Protease inhibitor, by Solarbio (1 mentions)
Anti acc, by Cell Signaling Technology (1 mentions)
Anti oxphos, by Abcam (1 mentions)
Ab110413, by ABclonal (1 mentions)
Anti sirt1, by Abcam (1 mentions)
Ripa lysis buffer, by Fdbio (1 mentions)
Anti ampk, by Cell Signaling Technology (1 mentions)
4 protocols using anti cd36
1
Western Blot Analysis of Key Proteins
2
Western Blotting for Protein Expression Analysis
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Western blotting analysis was performed as described in the previous report (Zhao et al., 2021 (link)). The cells were lysed in lysis buffer. After centrifugation, the supernatant was collected and the protein concentration was quantified by BCA assay kit. The samples were boiled at 95°C for 5 min and immediately frozen in a –80°C refrigerator. The primary antibodies were used as follows: anti-CD36 (ABclonal, Wuhan, China, A5792, 1:1,000), anti-GAPDH (ABclonal, Wuhan, China, AC033, 1:1,000), anti-FTO (ABclonal, Wuhan, China, A1438, 1:1,000), anti-P65-S536 (ABclonal, Wuhan, China, AP0475, 1:1,000), anti-IKB-α (Proteintech, Wuhan, China, 10268-1-AP, 1: 1,000), and anti-β-actin (ABclonal, Wuhan, China, AC026, 1: 1,000).
3
Protein Expression Analysis in Tissue Lysate
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Whole tissue lysate was prepared with RIPA buffer supplemented with phosphatase inhibitor and protease inhibitor cocktail before the experiment. Western blotting was performed by utilizing a standard protocol as described previously [23]. The antibodies were anti-β-actin, anti-PPARγ, anti- NF-κB/phospho-NF-κB, anti- eIF2α/phospho-eIF2α, anti-AKT/phospho-AKT (Ser-473), anti- GSK-3β/phosphor-GSK-3β (Cell Signaling Technology, Beverly, MA, USA), anti- SCD1, anti-CD36 (ABclonal Technology, Woburn, MA, USA) and anti-RNF186 (Sangon Biotech, Shanghai, China).
4
Western Blot Analysis of Metabolic Proteins
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Differentiated primary satellite cells in 6- or 12-well plates were quickly lysed in situ in 1x SDS-sample buffer (200 μl/well for 6-well plates or 100 μl/well for 12-well plates), followed by denaturation. For muscle tissue samples, frozen samples (20 mg) were immediately ground on ice in 200 μl RIPA lysis buffer (Fdbio science, #FD008) in the presence of protease inhibitors (Solarbio, P0100). Samples were then homogenized in 5× SDS-sample buffer at a final concentration of 1×, followed by denaturation. Protein samples were run on a 10% SDS-polyacrylamide gel, and Western blot analysis was performed. Primary antibodies were diluted 1:1000, and included anti-phosphorylated (p) AMPK (Cell signaling technology 2535), anti-AMPK (Cell signaling technology 2532), anti-phosphorylated (p) ACC (Cell signaling technology 3661 L), anti-ACC (Cell signaling technology 3662), anti-SIRT1 (Abcam, ab110304), anti-CPT1b (Proteintech, 22,170–1-AP), anti-CD36 (Abclonal, A19016), anti-PGC1a (Abclonal, A12348), anti-OXPHOS (Abcam, ab110413) and anti-GAPDH (Abclonal, Ac001). GAPDH was used as a loading control. The bands on Western blots were analyzed using Image J.
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