This is a knockout-validated antibody summary, based on the publication "Sirt1 regulates glial progenitor proliferation and regeneration in white matter after neonatal brain injury", as cited below [1], and "Expression of human inducible nitric oxide synthase in response to cytokines is regulated by hypoxia-inducible factor-1" for western blot knockout validation (figure 5b) [2]. 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 of HIF1a expression in white matter of Hif1a KO mice. Actin was used as loading control. From [1].

Mouse monoclonal IgG2b

Company: Novus

Antibody: HIF-1 alpha

Catalog number: NB100-123

Summary: Mouse monoclonal IgG2b against a fusion protein containing amino acids 432-528 of human HIF-1 alpha. Reacts with human, mouse, rat, porcine, avian, bovine, canine, ferret, primate and sheep. Suitable for western blot, immunoprecipitation, ChIP, immunocytochemistry/immunofluorescence, immunohistochemistry, flow cytometry, gel super shift assay and ELISA.

Western blot

White matter brain samples from WT and Hif1a KO mice. Tissues were homogenized in RIPA lysis buffer with proteinase inhibitors (Santa Cruz Biotech, Inc., Sc 24948).

5% milk in TBST for 1 h.

At 4 C overnight.

Appropriate horseradish peroxidase-conjugated secondary antibodies.

Chemiluminescent ECL substrate (Amersham, RPN2132) and visualized by exposure to X-ray-sensitive film.

Polyclonal Sirt1 antibody (1:250; Cell Signaling, 2028) is also KO validated by IC (Figure 5a).

The same clone (H1alpha67) is sold as Novus Biologicals NB100-105, NB100-123, NBP2-80760; OriGene TA309691; LifeSpan Biosciences LS-C2557, LS-B110, LS-C87583, LS-C87584, LS-C140776, LS-C140777, LS-C140778, LS-C140780, LS-C140802, LS-C140803, LS-C140804, LS-C140805, LS-C140806; Abnova MAB1892; Abcam ab1; Invitrogen MA1-16504, MA1-16511, MA1-46457, MA5-16008, MA5-16010, MA5-16009.