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2 protocols using abca1 antibody

1

Cholesterol Efflux Assay Protocol

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RPMI 1640 medium and fetal bovine serum (FBS) were purchased from Gibco (New York, NY, USA). Hcy, phorbol-12-myristate-13-acetate (PMA) and Oil-red O dye were supplied from Sigma-Aldrich (St. Louis, MO, USA). SBC-115076 and T0901317 was purchased from Selleck Chemicals (Houston, TX, USA). PCSK9 antibody was obtained from Cell Signaling Technology (Danvers, MA, USA). LDLR, ABCG1, CD68, β-actin and horseradish peroxidase (HRP) goat-anti-rabbit antibodies were both obtained from Abcam (Cambridge, MA, USA). ABCA1 antibody was purchased from Novus Biologicals (Littleton, CO, USA). LXRα was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). 22- [- (7- nitrobenz- 2- oxa- 1,3- diazol- 4- yl) amino]- 23, 24- bisnor- 5- cholen- 3β- ol (22-NBD cholesterol) was obtained from Invitrogen (Grand Island, NY, USA). PCR primers were supplied from Sangon Biotech (Shanghai). ApoA-I was supplied from Calbiochem (San Diego, CA, USA) and oxidized-LDL and high-density lipoprotein (HDL) were both obtained from Xiesheng Biotechnology (Beijing).
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2

Adipocyte Lipid Accumulation Assay

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Rosiglitazone and T0901317 were from Cayman Chemical (Ann Arbor, MI). Bexarotene was from LC Laboratories (Woburn, MA). The perilipin antibody was from Cell Signaling Technology (Danvers, MA). The ABCA1 antibody was from Novus Biologicals (Littleton, CO). Human insulin, Nile Red, p-nitrophenyl phosphate (pNPP) reagent, LG100268 and tributyltin chloride were from Sigma-Aldrich (St. Louis, MO). HX531 and T0070907 were from Tocris Bioscience (via R&D Systems, Inc., Minneapolis, MN). All other reagents were from Thermo Fisher Scientific (Suwanee, GA) unless noted. Caution: Tributyltin is hazardous and should be handled carefully.
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