Proteins from tissues or cells were harvested using radioimmunoprecipitation buffer containing Tris‐HCl (50 mmol L−1), NaCl (150 mmol L−1), MgCl2 (5 mmol L−1), EDTA (2 mmol L−1), NaF (1 mmol L−1), 1% NP40, and 0.1% SDS. The protein concentrations were quantified using commercial kits (Pierce). All protein samples were equally subjected to 10% SDS polyacrylamide gels, transferred to polyvinylidene difluoride membranes by electrophoresis, incubated with primary and secondary antibodies, and finally visualized by a chemiluminescence detection kit (Millipore). The following primary antibodies were purchased: NR2F6 (Abcam, Catalog number: ab137496; R&D Company, Catalog number: PP‐N2025‐00), CD36 (Abcam, Catalog number: ab133625), GAPDH (Abcam, Catalog number: ab181602), phosphorylated P65 (Beyotime, Catalog number: AF5881), total P65 (Beyotime, Catalog number: AF0246), a‐Tubulin (Sigma‐Aldrich, Catalog number: T6199).
Nr2f6
Manufactured by Abcam
NR2F6 is a nuclear receptor protein that functions as a transcriptional regulator. It is involved in the regulation of gene expression.
Automatically generated - may contain errors
Lab products found in correlation
Chemiluminescence detection kit, by Merck Group (1 mentions)
Gapdh, by Abcam (1 mentions)
Phosphorylated p65, by Beyotime (1 mentions)
Imagequant las 4000, by GE Healthcare (1 mentions)
Polyvinylidene difluoride membrane, by Merck Group (1 mentions)
Hrp conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions)
Flag tag (flag), by Merck Group (1 mentions)
Pgc 1α, by Proteintech (1 mentions)
C ebpα, by Santa Cruz Biotechnology (1 mentions)
Pparγ, by Cell Signaling Technology (1 mentions)
Las 4000, by Cytiva (1 mentions)
2 protocols using nr2f6
1
Protein Isolation and Western Blotting
2
Immunoblot Analysis of Protein Expression
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The tissues or cells were lysed and denatured at 100 °C for 10 min. The protein concentration was determined using the bicinchoninic acid assay. Equal amounts of protein were separated by SDS-PAGE, transferred to a polyvinylidene difluoride membrane (Millipore Corp), and immunoblotted overnight at 4 °C with the following primary antibodies: NR2F6 (Abcam; ab137496), PPARγ (Cell Signaling Technology; 81B8), PGC-1α (Proteintech; 66369-1-Ig), Flag tag (Sigma–Aldrich; F1804), UCP1 (Abcam; ab10983), C/EBPα (Santa Cruz; sc-365318) (422/aP2 antibody was kindly provided from M. Daniel Lane). After washing three times, the membranes were further incubated with the corresponding HRP-conjugated secondary antibodies (Jackson ImmunoResearch) for 1 h at room temperature. Protein bands were visualized using a chemiluminescence detection kit (ShareBio, Cat# SB-WB012) and analyzed with a luminescent ImageQuant LAS 4000 image analyzer (GE Healthcare Life Sciences).
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