Application

Research CategoryApoptosis & Cancer

Detect HAF/SART1 using this rabbit polyclonal Anti-HAF/SART1, Cat. No. ABC1431, validated for use in Chromatin Immunoprecipitation (ChIP), Immunoprecipitation, and Western Blotting.

Chromatin Immunoprecipitation (ChIP) Analysis: A representative lot detected HAF occupancy at the hypoxia response element (HRE) within the VEGFA and OCT-3/4 promoter region using chromatin preparation from human renal carcinoma 768-O (786-0) cells (Koh, M.Y. et al. (2015). Cancer Res. 75(2):316-329).

Immunoprecipitation Analysis: A representative lot immunoprecipitated HAF from human renal carcinoma ACHN cell lysate (Koh, M.Y. et al. (2015). Cancer Res. 75(2):316-329).

Western Blotting Analysis: Representative lots detected both endogenous and exogenously expressed HAF in various transfected human cancer cells, while a downregulated HAF was detected in cells co-transfected with HAF siRNA (Koh, M.Y. et al. (2015). Cancer Res. 75(2):316-329; Koh, M.Y. et al. (2011). Cancer Res. 71(11):4015-4027; Koh, M.Y., et al. (2008). Mol. Cell. Biol. 28(23):7081-7095).

Western Blotting Analysis: A representative lot detected HAP in HIF-1 immunoprecipitate from A549, HCT-116, and HT-29 xenografts (Koh, M.Y., et al. (2008). Mol. Cell. Biol. 28(23):7081-7095).

Disclaimer

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.

General description

U4/U6.U5 tri-snRNP-associated protein 1 (UniProt O43290; also known as Allergen Hom s 1, HAF, hSART-1, hSnu66, Hypoxia-associated factor, SART-1, SNU66 homolog, Squamous cell carcinoma antigen recognized by T-cells 1, U4/U6.U5 tri-snRNP-associated 110 kDa protein) is encoded by the SART1 (also known as HAF) gene (Gene ID 9092) in human. The hypoxia-associated factor (HAF) is an E3 ubiquitin ligase that mediates the ubiquitination and degradation of hypoxia-inducible factor 1alpha (HIF-1alpha). HAF also binds to HIF-2alpha and promotes HIF-2 transactivation instead of promoting HIF-2alpha degradation. In addition, hypoxia-dependent HAF SUMOylation enhances HAF interaction with HIF-2alpha and induction of HIF-2 target genes transcription, such as OCT-3/4, SOX2 and NANOG. HAF therefore promotes stem cell characteristics, aggressive tumor growth and invasion by switching cells from HIF-1alpha- to HIF-2alpha-dependent signaling. HAF overexpression enhances glioblastoma cells that express both HIF-1 and HIF-2alpha to initiate tumors as intracranial xenografts in mice. On the other hand, HAF overexpression results in decreased xenograft tumor growth of HT29 colon carcinoma cells that only express HIF-1alpha.

Immunogen

GST-tagged recombinant human HAF/SART1 C-terminal fragment.

Other Notes

Concentration: Please refer to lot specific datasheet.

Physical form

Unpurified.

Rabbit polyclonal antibody serum with 0.05% sodium azide.

Quality

Evaluated by Western Blotting in U2OS nuclear extract.

Western Blotting Analysis: A 1:2,000 dilution of this antiserum detected HAF/SART1 in 10 µg of U2OS nuclear extract.

Specificity

Target specificity was confirmed by Western blotting analysis of HAF overexpression and HAF shRNA-mediated HAF knockdown in various human cancer cells (Koh, M.Y., et al. (2008). Mol. Cell. Biol. 28(23):7081-7095).

Storage and Stability

Stable for 1 year at -20°C from date of receipt.

Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Target description

~120 kDa observed. 90.26 kDa calculated. The larger-than-calculated target band size is consistent with that reported in the literature (Koh, M.Y. et al. (2015). Cancer Res. 75(2):316-329). Uncharacterized bands may be observed in some lysate(s).