SREBP-2 Transcription Factor Assay Kit (ab133111)
Key features and details
- Assay type: Semi-quantitative
- Detection method: Colorimetric
- Platform: Microplate reader
- Sample type: Adherent cells, Nuclear Extracts, Suspension cells
Overview
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Product name
SREBP-2 Transcription Factor Assay Kit -
Detection method
Colorimetric -
Sample type
Adherent cells, Suspension cells, Nuclear Extracts -
Assay type
Semi-quantitative -
Species reactivity
Reacts with: Human
Predicted to work with: Mammals
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Product overview
Abcam's SREBP-2 Transcription Factor Assay Kit (ab133111) 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 a specific primary antibody directed against SREBP. A secondary antibody conjugated to HRP is added to provide a sensitive colorimetric readout at 450 nm.
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Notes
Lipid homeostasis in vertebrate cells is regulated by a family of transcription factors called sterol regulatory elements binding proteins (SREBPs). SREBPs directly activate the expression of over 30 gened involved in teh synthesis and uptake of cholesterol, fatty cids, triglycerides and phopholipids. SREBP-2 activates cholesterol synthesis by upregulating expression of HMG-CoA reductase.
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Platform
Microplate reader
Properties
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Storage instructions
Please refer to protocols. -
Components 96 tests 96-Well Plate Cover 1 unit Polysorbate 20 1 vial Transcription Factor Antibody Binding Buffer (10X) 1 x 3ml Transcription Factor Binding Assay Buffer (4X) 1 x 3ml Transcription Factor Developing Solution 1 x 12ml Transcription Factor Goat Anti-Rabbit HRP Conjugate 1 x 100µl Transcription Factor Reagent A 1 x 120µl Transcription Factor SREBP 96-Well Strip Plate 1 unit Transcription Factor SREBP Competitor dsDNA 1 vial Transcription Factor SREBP-2 Positive Control 1 vial Transcription Factor SREBP-2 Primary Antibody 1 vial Transcription Factor Stop Solution 1 x 12ml Wash Buffer Concentrate (400X) 1 x 5ml -
Research areas
- Epigenetics and Nuclear Signaling
- Transcription
- Domain Families
- HLH / Leucine Zipper
- HLH / Leucine Zipper
- Metabolism
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
- Lipid metabolism
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Function
Transcriptional activator required for lipid homeostasis. Regulates transcription of the LDL receptor gene as well as the cholesterol and to a lesser degree the fatty acid synthesis pathway (By similarity). Binds the sterol regulatory element 1 (SRE-1) (5'-ATCACCCCAC-3') found in the flanking region of the LDRL and HMG-CoA synthase genes. -
Tissue specificity
Ubiquitously expressed in adult and fetal tissues. -
Sequence similarities
Belongs to the SREBP family.
Contains 1 basic helix-loop-helix (bHLH) domain. -
Post-translational
modificationsAt low cholesterol the SCAP/SREBP complex is recruited into COPII vesicles for export from the ER. In the Golgi complex SREBPs are cleaved sequentially by site-1 and site-2 protease. The first cleavage by site-1 protease occurs within the luminal loop, the second cleavage by site-2 protease occurs within the first transmembrane domain and releases the transcription factor from the Golgi membrane. Apoptosis triggers cleavage by the cysteine proteases caspase-3 and caspase-7. -
Cellular localization
Nucleus and Endoplasmic reticulum membrane. Golgi apparatus membrane. Cytoplasmic vesicle > COPII-coated vesicle membrane. Moves from the endoplasmic reticulum to the Golgi in the absence of sterols. - Information by UniProt
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Alternative names
- AI608257
- bHLHd2
- Class D basic helix-loop-helix protein 2
see all -
Database links
- Entrez Gene: 6721 Human
- Omim: 600481 Human
- SwissProt: Q12772 Human
- Unigene: 443258 Human
Images
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ELISA ab133111 SREBP-2 Transcription Factor Assay Kit
HepG2 cells were grown in protein free hybridoma medium overnight (Life Technologies), followed by a 30 minutes incubation with the addition of 1% (2-Hydroxypropyl)-β-cyclodextrin (Sigma) or with the supplement of liqued cholesterol (Life Technologies). 10 uL of nuclear extracts (ab113474) were tested, corresponding to 2.5e5 cells per well.
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ELISA ab133111 SREBP-2 Transcription Factor Assay KitPositive control (10 or 5 uL) with or without inhibitor (10 or 2 uL) (duplicates; +/- SD).
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Functional Studies - SREBP-2 Transcription Factor Assay Kit (ab133111)Example of typical standard curve obtained using ab133111.
