SREBP Pathway

Membrane-bound transcription factors, sterol regulatory element binding protein-1 and -2 (SREBP-1 and SREBP-2) mediate homeostasis of cholesterol and fatty acids. SREBP-2 monitors cellular cholesterol levels and responds to low levels of cholesterol by the transcription of genes for HMG-CoA Reductase and other enzymes involved in cholesterol synthesis pathway. Recent studies implicate SREBP-2 in hypercholesterolemia, coronary heart disease, obesity, autophagy and Alzheimer's disease. Eurofins DiscoverX offers novel direct, functional cellular assays for SREBP2 translocation. No more assay development required; a simple mix-and-read microtiter plate-based assay is now available!


Features and Benefits
 

  • Whole cell based chemiluminescent assay
  • Non-imaging approach to monitor SREBP translocation
  • Simple instrumentation: Works on any standard luminometer


Applications
 

  • Novel, functional assay: For cholesterol sensing pathway
  • Screen or profile for SITE1 inhibitors
  • Measure effect of compounds on SREBP2 translocation


Assay List

Cell Lines

Target Description Cat. No.
SREBP2 PathHunter® U2OS SREBP2 Nuclear Translocation Cell Line 93-0830C3

Assay Ready Kits
Target Description Cat. No.
SREBP2 PathHunter® eXpress SREBP2 Nuclear Translocation Assay 93-0830E3CP5L
SREBP2 PathHunter® eXpress SREBP2 Nuclear Translocation Assay 93-0830E3CP5M
SREBP2 PathHunter® eXpress SREBP2 Nuclear Translocation Assay 93-0830E3CP5S


Assay Principle

PathHunter® cell lines feature novel in vivo applications of Enzyme Fragment Complementation (EFC) technology in which the β-galactosidase (β-gal) enzyme has been split into two inactive fragments ProLink™  and enzyme acceptor. DiscoveRx has designed an EFC assay to monitor nuclear translocation of SREBP2. A small 4 kDa complementing fragment of β-gal, ProLink™ (PK) tag has been fused recombinantly to the SREBP2 protein. The larger portion of β-gal, termed EA for ‘enzyme acceptor’, is anchored in the nucleus. Low cholesterol activates PK-SREB2/Scap protein complex, which blebs off from endoplasmic reticulum and traverses to Golgi where it is cleaved by S1P. The SREBP2-PK protein translocates to nucleus where it binds to EA resulting in a complemented enzyme. Enzyme Fragment Complementation is measured quantitatively using the chemiluminescent substrate in the PathHunter®  Detection Kit.

How it Works

 

PathHunter® SREBP2 Nuclear Translocation Assay