This is a knockout-validated antibody summary, based on the publication "The metabolome regulates the epigenetic landscape during naive-to-primed human embryonic stem cell transition", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

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Figure 1. Western blot analysis of HIF1α expression in cells pushed toward the primed stage by culture in TeSR1 for 5 days in wild type Elf1 cells (iCas9 Elf1), and two CRISPR-Cas9 KO mutants of HIF1α (gHIF1 6.2.1 and gHIF1 6.3.1). Tubulin was used as loading control. From [1].

Mouse monoclonal IgG1, κ

Company: BD Biosciences

Antibody: HIF-1

Catalog number: 610958

Summary: Mouse monoclonal IgG1, κ against human HIF-1α aa. 610-727. Reacts with human by western blot and immunofluorescence.

Western blot

Wild type and HIF1α KO Elf1 cells. Cellular extracts were prepared using a lysis buffer containing 20 mM Tris HCl (pH 7.5), 150 mM NaCl, 15% glycerol, 1% Triton, 25mM β-glycerophosphate, 50mM NaF, 10mM Na Pyrophosphate, orthovanadate, PMSF (all chemicals are from Sigma-Aldrich, St. Louis, MO), Protease inhibitor cocktail (Roche) and 2% SDS.

5% nonfat dry milk for 60 minutes at RT.

1/2,000 dilution overnight at 4°C.

Horseradish peroxidase-conjugated secondary antibodies for 1 h.

Enhanced chemiluminescence (Millipore Corp, Billerica, MA).