HIF-1 alpha [Hydroxy Pro564] Antibody - BSA Free
Novus Biologicals, part of Bio-Techne | Catalog # NB110-74679
Key Product Details
Validated by
Biological Validation
Species Reactivity
Validated:
Human, Mouse
Cited:
Human, Mouse
Applications
Validated:
ELISA, Immunoblotting, Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot
Cited:
IF/IHC, Immunoblotting, Western Blot
Label
Unconjugated
Antibody Source
Polyclonal Rabbit Serum
Format
BSA Free
Concentration
This product is unpurified. The exact concentration of antibody is not quantifiable.
Product Specifications
Immunogen
This HIF-1 alpha [Hydroxy Pro564] Antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to a region surrounding the P564 of human HIF-1 alpha. (Uniprot: Q16665)
Reactivity Notes
A BLAST analysis was used to suggest cross-reactivity with HIF-1 alpha from monkey, rat, dog, bovine and Xenopus sources based on a 100% homology with the immunizing sequence. Reactivity against homologues from other sources is not known
Modification
Hydroxy Pro564
Localization
Cytoplasmic in normoxia, nuclear translocation in response to hypoxia.
Specificity
This antibody is directed against human HIF-1a hydroxyP564 and is specific for the hydroxylated form of the protein. Minimal reactivity occurs with the non-hydroxylated form of the protein. This antibody is specific for HIF-1a hydroxylated at P564. Minimal cross-reactivity occurs with non-hydroxylated HIF-1a. A BLAST analysis was used to suggest cross-reactivity with HIF-1a from human, monkey, mouse, rat, dog, bovine and Xenopus sources based on a 100% homology with the immunizing sequence. Reactivity against homologues from other sources is not known.
Clonality
Polyclonal
Host
Rabbit
Isotype
Serum
Description
This antibody is directed against human HIF-1a hydroxy P564 and is specific for the hydroxylated form of the protein
Store vial at -20C prior to opening. Aliquot contents and freeze at -20C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use.
Store vial at -20C prior to opening. Aliquot contents and freeze at -20C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use.
Scientific Data Images for HIF-1 alpha [Hydroxy Pro564] Antibody - BSA Free
Western Blot: HIF-1 alpha [Hydroxy Pro564] Antibody [NB110-74679] - Detection (left panel) of HIF-1 alpha [Hydroxy Pro564] in nuclear extracts of A549 cells treated with MG132 (a proteasome inhibitor). Hydroxyproline is not recognized on HIF-1alpha when cells are first treated with DFO, a prolyl hydroxylase inhibitor that prevents HIF hydroxylation. Control staining is shown (right panel) using conventional anti-HIF-1alpha. The asterisk marks a band approximately 110 kDa molecular weight in size corresponding to HIF-1 alpha. The primary antibody was used at a 1:1000 dilution in 2% BLOTTO.
Applications for HIF-1 alpha [Hydroxy Pro564] Antibody - BSA Free
Application
Recommended Usage
ELISA
1:5000-1:20000
Immunoblotting
1:100-1:2000
Immunohistochemistry
1:10 - 1:500
Immunohistochemistry-Paraffin
1:10 - 1:500
Western Blot
1:500-1:2000
Application Notes
This antibody has been tested for use in ELISA and western blotting. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 110 kDa in size corresponding to HIF-1a hydroxyl P564 by western blotting in the appropriate cell lysate or extract.
Use in Immunoblotting reported in scientific literature (PMID 28566998).
Use in Immunoblotting reported in scientific literature (PMID 28566998).
Reviewed Applications
Read 3 reviews rated 4 using NB110-74679 in the following applications:
Formulation, Preparation, and Storage
Purification
Unpurified
Formulation
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Format
BSA Free
Preservative
0.01% Sodium Azide
Concentration
This product is unpurified. The exact concentration of antibody is not quantifiable.
Shipping
The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.
Background: HIF-1 alpha/HIF1A
HIF-1 or hypoxia inducible factor 1 (predicted molecular weight 93kDa), is a transcription factor commonly referred to as a "master regulator of the hypoxic response" for its central role in the regulation of cellular adaptations to hypoxia. In its active form under hypoxic conditions, HIF-1 is stabilized by the formation of a heterodimer of HIF-1 alpha and ARNT/HIF-1 beta subunits. Nuclear HIF-1 engages p300/CBP for binding to hypoxic response elements (HREs). This process induces transcription and regulation of genes including EPO, VEGF, iNOS2, ANGPT1 and OCT4 (4,5).
Under normoxic conditions, the HIF-1 alpha subunit is rapidly targeted and degraded by the ubiquitin proteasome system. This process is mediated by prolyl hydroxylase domain enzymes (PHDs), which catalyze the hydroxylation of key proline residues (Pro-402 and Pro-564) within the oxygen-dependent degradation domain of HIF-1 alpha. Once hydroxylated, HIF-1 alpha binds the von Hippel-Lindau tumor suppressor protein (pVHL) for subsequent ubiquitination and proteasomal degradation (4). pVHL dependent regulation of HIF-1 alpha plays a role in normal physiology and disease states. Regulation of HIF-1 alpha by pVHL is critical for the suppressive function of FoxP3+ regulatory Tcells (6). Repression of pVHL expression in chronic lymphocytic leukemia (CLL) B cells leads to HIF-1 alpha stabilization and increased VEGF secretion (7).
References
1. Semenza, G. L., Agani, F., Feldser, D., Iyer, N., Kotch, L., Laughner, E., & Yu, A. (2000). Hypoxia, HIF-1, and the pathophysiology of common human diseases. Advances in Experimental Medicine and Biology.
2. Muz, B., de la Puente, P., Azab, F., & Azab, A. K. (2015). The role of hypoxia in cancer progression, angiogenesis, metastasis, and resistance to therapy. Hypoxia. https://doi.org/10.2147/hp.s93413
3. Huang, Y., Lin, D., & Taniguchi, C. M. (2017). Hypoxia inducible factor (HIF) in the tumor microenvironment: friend or foe? Science China Life Sciences. https://doi.org/10.1007/s11427-017-9178-y
4. Koyasu, S., Kobayashi, M., Goto, Y., Hiraoka, M., & Harada, H. (2018). Regulatory mechanisms of hypoxia-inducible factor 1 activity: Two decades of knowledge. Cancer Science. https://doi.org/10.1111/cas.13483
5. Dengler, V. L., Galbraith, M. D., & Espinosa, J. M. (2014). Transcriptional regulation by hypoxia inducible factors. Critical Reviews in Biochemistry and Molecular Biology. https://doi.org/10.3109/10409238.2013.838205
6. Lee, J. H., Elly, C., Park, Y., & Liu, Y. C. (2015). E3Ubiquitin Ligase VHL Regulates Hypoxia-Inducible Factor-1 alpha to Maintain Regulatory T Cell Stability and Suppressive Capacity. Immunity. https://doi.org/10.1016/j.immuni.2015.05.016
7. Ghosh, A. K., Shanafelt, T. D., Cimmino, A., Taccioli, C., Volinia, S., Liu, C. G., ... Kay, N. E. (2009). Aberrant regulation of pVHL levels by microRNA promotes the HIF/VEGF axis in CLL B cells. Blood. https://doi.org/10.1182/blood-2008-10-185686
Long Name
Hypoxia Inducible Factor 1 Subunit Alpha
Alternate Names
BHLHE78, HIF 1A, HIF-1a, HIF1 alpha, HIF1A, MOP1, PASD8
Entrez Gene IDs
3091 (Human)
Gene Symbol
HIF1A
UniProt
Additional HIF-1 alpha/HIF1A Products
Product Documents for HIF-1 alpha [Hydroxy Pro564] Antibody - BSA Free
Product Specific Notices for HIF-1 alpha [Hydroxy Pro564] Antibody - BSA Free
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
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