Anti-IspH

4-hydroxy-3-methylbut-2-enyl diphosphate reductase (UniProt: P62623; also known as EC:1.17.7.4, HMBPP reductase, 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase) is encoded by the ispH (also known as lytB, yaaE, b0029, JW0027) gene (Gene ID: 944777) in E. coli. IspH is an enzyme that catalyzes the conversion of 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate (HMBPP) into a mixture of isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). It contains a central iron-sulfur cluster that serves as an electron transfer cofactor in the active site. IspH acts in the terminal step of the methyl erythritol phosphate (MEP) pathway for isoprenoid precursor biosynthesis that is essential for survival of Gram-negative bacteria, mycobacteria, and apicomplexans. It operates at an optimum pH of 7.0. Its substrate binding site is localized in amino acids 225-227. Since MEP pathway is absent in human, inhibitors of IspH are attractive targets for the development of antibiotics. More recently, ester-based prodrug of C23 analog have been described that inhibit IspH and are highly potent against Gram-negative bacteria. (Ref.: Singh, KS., et al. (2021). Nature 589(7843); 597-602; Grawert, T., et al. (2004). J. Am. Chem. Soc. 126(40); 12847-12855).

This rabbit polyclonal antibody specifically detects 4-hydroxy-3-methylbut-2-enyl diphosphate reductase (IspH) in E.coli.

Purified E. coli IspH protein was mixed with the KLH conjugate and Freud s complete adjuvant.

Quality Control Testing

Evaluated by Western Blotting in E. coli lysates of strain CGSC8074 (positive) with conditional knockdown of IspH.

Western Blotting Analysis: A 1:5,000 dilution of this antibody detected IspH in lysate from E. coli, strain CGSC8074, but not in lysate from E.coli with conditional knockdown of IspH.

Tested Applications

Western Blotting Analysis: A representative lot detected IspH in Western Blotting applications (Singh, K., et al. (2021). Nature. 589(7843):597-602).

Enzyme Immunoassay Analysis (ELISA): A representative lot detected IspH in ELISA applications (Singh, K., et al. (2021). Nature. 589(7843):597-602).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user

Anti-IspH, Cat. No. ABF998, is a rabbit polyclonal antibody that detects 4-hydroxy-3-methylbut-2-enyl diphosphate reductase (IspH) and is tested for use in ELISA and Western Blotting.

Purified rabbit polyclonal antibody in PBS with 0.02% sodium azide.

Recommend storage at +2°C to +8°C. For long term storage antibodies can be kept at -20°C. Avoid repeated freeze-thaws.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

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.