Anti-DHFR, N-terminal antibody produced in rabbit

Anti-DHFR, N-terminal antibody produced in rabbit has been used in immunoprecipitation and western blotting.

Anti-DHFR, N-terminal antibody is specific for the epitope residing within amino acids of mouse DHFR. The antibody interacts with DHFR and DHFR fusion proteins. The product is specific for DHFR in humans, mice and rats. Staining of the DHFR band by immunoblotting is inhibited by the immunizing peptide.

Dihydrofolate reductase (DHFR) inhibitors, such as methotrexate, are folate analogs which can bind to the active site and deactivate the enzyme. From another angle, structural and enzymatic properties of DHFR led to development of a variety of screenings, in which DHFR can function in a fashion that resembles reporter genes. The screenings are based on the fact that DHFR can be dissected into two halves that can reassemble to form an active enzyme. Thus, each of the two halves of DHFR can be expressed as two fusion proteins that when interacting with each other can restore the DHFR enzymatic activity. The readout of this protein-protein interaction and consequent enzymatic activity, can be either restoration of growth in bacterial, yeast and plant, or receptor activation. The approach was proved to be useful for receptor-ligand, antigen-antibody and other interactions.

Dihydrofolate reductase (DHFR) is an NADPH dependent enzyme that reduces dihydrofolate and regenerates tetrahydrofolate. DHFR is involved in the biosynthesis of thymidylate and purines, and thereby regulates DNA synthesis and cell survival. Alterations in DHFR have been associated with impaired nucleic acid synthesis and cell death. Thus, this enzyme has important therapeutic implications in cell proliferative disorders such as cancer.

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Dihydrofolate reductase has 187 amino acids and corresponds to molecular weight of 18-20 kDa. It is mapped to chromosome 5q22 region. DHFR contains substrate binding site and coenzyme binding domain.

The immunizing sequence is conserved in mouse, human and rat.

Solution in 0.01 M phosphate buffered saline, pH 7.4, containing 1% bovine serum albumin and 15 mM sodium azide.