Tissues were homogenized and lysed in lysis buffer as previously described [40 (link)]. The protein concentration was determined by Bradford protein assay (Bio-Rad). Equal amounts of proteins were loaded and immunoblot analyses were carried out according to standard protocol. The following antibodies from Cell Signaling Technologies were used: HSL (4107), Phospho-HSL (Ser563) (4139), ATGL (2138), HSP60 (12165), DRP1 (5391), phosphor-DRP1 Ser616 (4494), phosphor-DRP1 Ser637 (4867), phosphor-AKT Ser473 (4060), AKT (9272), and LC3A/B (4108). Total OXPHOS Rodent WB Antibody Cocktail (ab110413), MLKL (ab172868), Anti-RIP3 (phosphor T231 + S232) (ab201912), and UCP1 (ab1426) were obtained from Abcam. GAPDH (60004-1-IG) and P62 (18420-1-AP) were obtained from Proteintech; PLIN1 (GP29, Progen Biotechnik GmbH); β-actin (MAB1501, MilliporeSigma); RIPK3 (NBP1-77299SS, Novus Biologicals).
Ab1426
Manufactured by Abcam
Ab1426 is a laboratory equipment product. It serves a core function related to the research and analysis process, though a detailed description cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Bradford protein assay, by Bio-Rad (1 mentions)
Phospho hsl ser563, by Cell Signaling Technology (1 mentions)
Gapdh 60004 1 ig, by Proteintech (1 mentions)
P62 18420 1 ap, by Proteintech (1 mentions)
Plin1 gp29, by Progen Biotechnik (1 mentions)
Fv1000 confocal fluorescence microscope, by Olympus (1 mentions)
Ab6640, by Abcam (1 mentions)
Ab59685, by Abcam (1 mentions)
2 protocols using ab1426
1
Comprehensive Protein Immunoblot Analysis
2
Immunofluorescence Imaging of Adipocyte Markers
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Cells were grown on glass coverslips and fixed with 4% paraformaldehyde in PBS (pH 7.4) for 20 min at 4 °C. Cells were permeabilized with 0.3% Triton-X 100 in PBS for 30 min and blocked with PBS containing 3% goat serum and 1% bovine serum albumin for 1 h at room temperature. After incubation with rabbit anti-UCP-1 (EMD Millipore, #AB1426), Adrb3 (Abcam, #ab59685), and rat anti-F4/80 (Abcam, #ab6640) at 1:100 dilution overnight at 4 °C, the cells were washed with PBS and treated with goat anti-rabbit or anti-rat Alexa Fluor 488-conjugated and Alexa Fluor 594-conjugated secondary antibodies at 1:1000 dilution for 1 h. Cells were washed with PBS and incubated in 0.1 μg/ml Hochest (DAPI) for 30 min. After washing with PBS, coverslips were mounted onto microscope slides. Images were recorded using an Olympus FV1000 confocal fluorescence microscope.
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