Novus Biologicals, a leading supplier of hypoxia-inducible factor (HIF) antibodies, recently launched two new HIF related antibodies: an anti-Factor Inhibiting HIF-1 antibody (catalog number NBP1-30333) and an anti-HIF Prolyl Hydroxylase 2 antibody (catalog number NBP1-30328). These two new antibodies complement Novus’ current line of hypoxia antibodies (~2,000). Read on to learn more about these two antibodies and HIF dynamics.

Novus’ newly released mouse monoclonal anti-Factor Inhibiting HIF-1 antibody (clone 162C) was made to the full length human Factor Inhibiting HIF-1 protein expressed in E. coli BL21 (DE3) cells. It is protein G purified and was QC tested in Western blot on A431 cell lysates, which resulted in a clear band at 40kDa. This antibody is also useful for immunohistochemistry on paraffin-embedded sections.

Novus’ newly released mouse monoclonal anti-HIF Prolyl Hydroxylase 2 antibody (clone 366G/76/3) was made to a peptide within residues 1-50 of the human protein. It is also protein G purified and was QC tested in Western blot on HeLa whole cell extracts, which resulted in a clear band at approximately 46kDa. As with the FIH antibody, this antibody is also useful for immunohistochemistry on paraffin-embedded sections.

These two antibodies are closely related to other HIF antibodies and hypoxia research. The Factor Inhibiting HIF-1 (FIH-1) acts as a transcriptional corepressor of HIF-1 transcriptional activity, by binding to VHL. By recruiting histone deacetylases, VHL inhibits transactivation caused by HIF-1 alpha. FIH-1 hydroxylates HIF-1 alpha at Asp(803) in the C-terminal transactivation domain (CAD), and under normoxic conditions, it acts as an oxygen sensor. HIF prolyl hydroxylase 2 (gene name EGLN1) is a prolyl hydroxylase that modifies HIF-alpha proteins; it hydroxylates HIF-1 alpha at Pro(402) and Pro(564), and HIF-2 alpha. Similar to FIH-1, EGLN1 is an oxygen sensor. Under the appropriate conditions, EGLN1 targets HIF for proteasomal degradation by the VHL ubiquitination complex.