Lipid homeostasis in vertebrate cells is regulated by a family of transcription factors called sterol regulatory element-binding proteins (SREBP’s). SREBP’s directly activate the expression of over 30 genes involved in the synthesis and uptake of cholesterol, fatty acids, triglycerides, and phospholipids.1 Three different SREBP isoforms designated SREBP-1a, SREBP-1c, and SREBP-2 are encoded by two different genes and belong to the basic helix-loop-helix-leucine zipper (bHLH-ZIP) family of transcription factors.2 SREBP-2 performs a critical role in the transcriptional regulation of genes involved in cholesterol synthesis and uptake including HMG-CoA synthase, HMG-CoA reductase, and the LDL receptor. Cayman’s SREBP-2 Transcription Factor Assay is a non-radioactive, sensitive method for detecting specific transcription factor DNA binding activity in nuclear extracts and whole cell lysates. A 96 well enzyme-linked immunosorbent assay (ELISA) replaces the cumbersome radioactive electrophoretic mobility shift assay (EMSA). A specific double stranded DNA (dsDNA) sequence containing the SREBP response element is immobilized to the wells of a 96 well plate. SREBP contained in a nuclear extract, binds specifically to the SREBP response element. SREBP is detected by addition of specific primary antibody directed against SREBP. A secondary antibody conjugated to HRP is added to provide a sensitive colorimetric readout at 450 nm.Needed but not supplied: Please download the kit booklet to verify if UltraPure Water (Milli-Q or equivalent) or any other components are needed for this assay.WARNING This product is not for human or veterinary use.
