SREBP-2 antibody

Overview


	Antigen:	SREBP-2
	Antibody Type:	Monoclonal
	Application:	Western Blotting (WB), Immunoprecipitation (IP)
	Host:	Mouse
	Quantity:	100 µg (1 mg/mL)
	Price:	191,00 € (Plus shipping costs and VAT)
	Order number:	ABIN131763
	Availability:	7 to 10 days until shipment

Hints on ordering

Add to basket

Customer Service

Technical Inquiry
Send product details to colleague
Download Datasheet

Additional Information: SREBP-2 antibody

Antigen	SREBP-2
Immunogen	This antibody was purified from hybridoma (clone 1D2) supernatant using protein A agarose. This hybridoma was established by fusion of mouse myeloma cell P3U1 with Balb/c mouse splenocyte immunized with human recombinant SREBP-2.
Antibody Type	Monoclonal
Isotype	IgG1 »Matching secondary antibodies
Description	Sterol regulatory binding proteins (SREBP) are membrane-bound transcription factors that regulate cholesterol and fatty acid homeostasis. Proteolytic cleavage of the 125 kDa SREBP by caspase-3 or SCA results in the N-terminal fragment entering the nucleus and activating transcription of enzymes involved in lipid synthesis. SREBP-1a and -1c are derived from a single gene via alternative transcription start sites while SREBP-2 is derived from a separate gene and is ~45% identical to SREBP-1a. SREBP-1 preferentially activates genes involved in fatty acid synthesis, whereas SREBP-2 preferentially activates genes involved in cholesterol biosynthesis such as hydroxymethylglutaryl (HMG) CoA synthase, HMG-CoA reductase, farnesyl diphosphate synthase and squalene synthase.
Clone	1D2
Host	Mouse
Specificity	This antibody reacts with human SREBP-2 (120 kDa) on Western blotting.

Application Details: SREBP-2 antibody

Application	Western Blotting (WB), Immunoprecipitation (IP)
Application Notes	SDS-PAGE & Western Blotting 1) Wash the cells 3 times with PBS and suspend with 10 volumes of cold Lysis buffer (50 mM Tris-HCl, pH 7.2, 250 mM NaCl, 0.1% NP-40, 2 mM EDTA, 10% glycerol) containing appropriate protease inhibitors. Incubate it at 4 o C with rotating for 30 minutes, then sonicate briefly (up to 10 seconds). 2) Centrifuge the tube at 12,000 x g for 10 minutes at 4 o C and transfer the supernatant to another tube. Measure the protein concentration of the supernatant and add the cold Lysis buffer to make an 8 mg/mL solution. 3) Mix the sample with equal volume of Laemmli’s sample buffer. 4) Boil the samples for 2 minutes and centrifuge. Load 10 µL of the sample per lane in a 1 mm thick SDS-polyacrylamide gel for electrophoresis. 5) Blot the protein to a polyvinylidene difluoride (PVDF) membrane at 1 mA/cm 2 for 1 hour in a semi-dry transfer system (Transfer Buffer - 25 mM Tris, 190 mM glycine, 20% MeOH). See the manufacture's manual for specific transfer procedure. 6) To reduce nonspecific binding, soak the membrane in 10% skimmed milk (in PBS, pH 7.2) for 1 hour at room temperature, or overnight at 4 o C. 7) Incubate the membrane with primary antibody diluted with PBS, pH 7.2 containing 1% skimmed milk as suggested in the Applications for 1 hour at room temperature. (The optimal antibody concentration will depend on the experimental conditions.) 8) Wash the membrane with PBS (5 minutes x 6 times). 9) Incubate the membrane with the 1:10,000 POD-conjugated anti-mouse IgG (MBL, code no. 330) diluted with 1% skimmed milk (in PBS, pH 7.2) for 1 hour at room temperature. 10) Wash the membrane with PBS (5 minutes x 6 times). 11) Wipe excess buffer from the membrane, then incubate it with appropriate chemiluminescence reagents for 1 minute. Remove extra reagent from the membrane by dabbing with a paper towel, and seal it in plastic wrap. 12) Expose to X-ray film in a dark room for 5 minutes. Develop the film as usual. The optimal conditions for exposure and development may vary. Positive controls for Western blotting, MCF-7, HEp-2, LNCaP 050816-1 Western blotting: 10 µg/mL for chemiluminescence detection system. Immunoprecipitation: Not recommended. Immunohistochemistry: Not recommended. Immunocytochemistry: Not tested. Flow Cytometry: Not tested. MBL was not satisfied with our results using this antibody for immunoprecipitation, so we do not recommend it for this application. However, the antibody has been successfully used for immunoprecipitation (3) in the scientific literature.
Concentration	1 mg/mL
Buffer	100 µg IgG in 100 µL volume of PBS containing 50% glycerol, pH 7.2. Contains no preservatives.
Restrictions	For Research Use Only

Publications: SREBP-2 antibody

Publications

Demoulin, Ericsson, Kallin et al.:
"Platelet-derived growth factor stimulates membrane lipid synthesis through activation of phosphatidylinositol 3-kinase and sterol regulatory element-binding proteins."
In: The Journal of biological chemistry , Vol. 279, Issue 34, pp. 35392-402, 2004/01 (PubMed)

Yabe, Brown, Goldstein:
"Insig-2, a second endoplasmic reticulum protein that binds SCAP and blocks export of sterol regulatory element-binding proteins."
In: Proceedings of the National Academy of Sciences of the United States of America , Vol. 99, Issue 20, pp. 12753-8, 2002/01 (PubMed)

Images: SREBP-2 antibody

Western blot analysis of SREBP-2 expression in HEp-2 cells (1), MCF-7 cells (2), LNCaP cells (3), HT1080 cells (4), ZR75-1 cells (5) and using ABIN131763.

Image for SREBP-2 antibody (1)

External Information: SREBP-2 antibody

Google Scholar	Find more references for clone 1D2 on Google Scholar™
EMBL Harvester	Search human, mouse or rat genome for antigen SREBP-2 (Bioinformatic Harvester (EMBL))