MABS1233

Anti-HMG-CoA Reductase Antibody, clone IgG-A9

clone IgG-A9, from mouse

• Synonym(s): 3-hydroxy-3-methylglutaryl-coenzyme A reductase, HMG-CoA reductase
• UNSPSC Code: 12352203
• eCl@ss: 32160702

Properties

• biological source: mouse
• Quality Level: 100
• antibody form: purified antibody
• antibody product type: primary antibodies
• clone: IgG-A9, monoclonal
• species reactivity: human, rat, hamster
• technique(s): immunocytochemistry: suitable; immunoprecipitation (IP): suitable; western blot: suitable
• isotype: IgG1κ
• UniProt accession no.: P00347
• shipped in: ambient
• target post-translational modification: unmodified
• Gene Information: human ... HMGCR(3156)

Description

General description

3-hydroxy-3-methylglutaryl-coenzyme A reductase (EC 1.1.1.34; UniProt P00347; also known as HMG-CoA reductase) is encoded by the HMGCR gene (Gene ID 100756363) in Cricetulus griseus (Chinese hamster). HMG-CoA reductase catalyzes the conversion of HMG-CoA to mevalonic acid, the rate-limiting enzyme of the mevalonate-isoprenoid biosynthesis (MIB) pathway responsible for the production of cholesterol and other isoprenoids, as well as farnesyl pyrophosphate (FPP) required for protein prenylation. HMG-CoA reductase is suppressed by cholesterol derived from the internalization and degradation of low density lipoprotein (LDL) via the LDL receptor as well as oxidized cholesterol species. Competitive inhibitors against HMG-CoA reductase induce LDL receptors expression in the liver, which in turn increases the catabolism of plasma LDL and lowers the plasma concentration of cholesterol. HMG-CoA reductase is a multitransmembrane ER protein with an N-terminal sterol-sensing domain (SSD; a.a. 61-218) and a C-terminal catalytic domain (a.a. 450-888) whose activity is inhibited upon SSD binding by cholesterol. Numerous studies have implicated the involvement of HMG-CoA reductase and mevalonate pathway in carcinogenesis.

Specificity

Clone IgG-A9 targets an epitope within HMC-CoA reductase catalytic domain (a.a. 450 887).

Immunogen

Membrane preapration from compactin-adapted CHO cells (Liscum, L., et al. (1983). J. Biol. Chem. 258(13) 8450-8455).

Application

Detect HMG CoA Reductase using this mouse monoclonal Anti-HMG-CoA Reductase, clone IgG-A9, Cat. No. MABS1233, validated for use in Immunocytochemistry, Immunoprecipitation, and Western Blotting.

Research Category
Signaling

Western Blotting Analysis: A 1:100 dilution from a representative lot detected in 15 µg of membrane extract from CHO7 cells treated overnight with 0.01 mM compactin (Courtesy of Linda Donnelly, Department of Molecular Genetics, UT Southwestern Medical Center, Dallas, TX, USA).

Immunocytochemistry Analysis: A representative lot detected HMG CoA reductase-positive ER membrane structures by indirect immunofluorescence staining of paraformaldehyde-fixed, 0.1% Triton X-100-permeabilized CHO-K1 cells cultured with compactin, mevalonate, and MG-132, in the presence or absence of 25-hydroxycholesterol (25-HC) (Hartman, I.Z., et al. (2010). J. Biol. Chem. 285(25):19288-19298).

Immunoprecipitation Analysis: A representative lot immunodepleted HMC-CoA reductase activity from compactin-adapted CHO cell (UT-1) extract (Liscum, L., et al. (1983). J. Biol. Chem. 258(13) 8450-8455).

Western Blotting Analysis: A representative lot detected 25-hydroxycholesterol (25-HC) treatment-induced HMC-CoA reductase dislocation from ER membrane to the cytosol and lipid droplet in CHO-K1 cells cultured with lipoprotein-deficient serum in the presence of compactin, MG-132, with or without mevalonate. Cellular VCP knockdown prevented sterol-induced HMC-CoA reductase cytosolic, but not lipid droplet, translocation (Hartman, I.Z., et al. (2010). J. Biol. Chem. 285(25):19288-19298).

Western Blotting Analysis: A representative lot detected 25-hydroxycholesterol (25-HC) treatment-induced ubiquitination of HMC-CoA reductase in SV-589 human fibroblasts cultured with compactin and mevalonate in the presence of proteasome inhibitor MG-132. Cellular knockdown of gp78A, but not gp78B or Hrd1, reduced 25-HC-induced HMC-CoA reductase ubiquitination (Song, B.L., et al. (2005). Mol. Cell. 19(6):829-840).

Western Blotting Analysis: A representative lot detected 25-hydroxycholesterol (25-HC) treatment-induced degradation of HMC-CoA reductase in SV-589 human fibroblasts cultured with compactin and mevalonate. Cellular knockdown of gp78A, VCP, or Insig-1/-2 suppressed sterol-induced HMC-CoA reductase degradation (Song, B.L., et al. (2005). Mol. Cell. 19(6):829-840).

Western Blotting Analysis: A representative lot detected HMC-CoA reductase in both membrane and nuclear fractions from HEK-293S cells cultured with compactin and mevalonate. Additional treatment with 25-hydroxycholesterol (25-HC) resulted in HMC-CoA reductase degradation (Sever, N., et al. (2003). Mol. Cell. 11(1):25-33).

Western Blotting Analysis: A representative lot detected an upregulated HMC-CoA reductase expression in compactin-adapted CHO cells (UT-1), as well as a loss of HMC-CoA reductase expression in UT-1 cells cultured in the presence of LDL (Liscum, L., et al. (1983). J. Biol. Chem. 258(13) 8450-8455).

Western Blotting Analysis: A representative lot detected a greater HMC-CoA reductase upregulation in liver microsome preparation from rats on diet supplemented with cholestyramine and mevinolin than with cholestyramine alone, while supplementation with cholesterol in addition to cholestyramine and mevinolin downregulated liver HMC-CoA reductase (Liscum, L., et al. (1983). J. Biol. Chem. 258(13) 8450-8455).

Quality

Identity Confirmation by Isotyping Test.

Isotyping Analysis: The identity of this monoclonal antibody is confirmed by isotyping test to be mouse IgG1 .

Target description

~92 kDa observed. 97.08 kDa (hamster), 97.48/92.02/99.75 kDa (human isoform 1/2/3 (HMGCR-1b)), 96.69 kDa (rat) calculated.. Uncharacterized bands may be observed in some lysate(s).

Physical form

Format: Purified

Protein G purified.

Purified mouse IgG1 in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt.

Other Notes

Concentration: Please refer to lot specific datasheet.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Safety Information

• Storage Class Code: 12 - Non Combustible Liquids
• WGK: WGK 1
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable