HIF-1 alpha Antibody
Novus Biologicals, part of Bio-Techne | Catalog # NB100-449
Key Product Details
Validated by
Knockout/Knockdown, Biological Validation
Species Reactivity
Validated:
Human, Mouse, Rat, Canine, Chicken, Goat, Monkey, Primate
Cited:
Human, Mouse, Rat, Avian - Chicken, Canine, Goat, Primate
Predicted:
Bovine (100%). Backed by our 100% Guarantee.
Applications
Validated:
Knockout Validated, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, ELISA, Flow Cytometry, Flow (Intracellular), Immunocytochemistry/ Immunofluorescence, Simple Western, Immunoprecipitation, Chromatin Immunoprecipitation (ChIP), Knockdown Validated
Cited:
Knockout Validated, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, ELISA, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Chemotaxis, Flow Cytometry Control, Proximity Ligation Assay, IF/IHC, Knockdown Validated
Label
Unconjugated
Antibody Source
Polyclonal Rabbit IgG
Concentration
0.2 mg/ml
Product Specifications
Immunogen
The immunogen recognized by this HIF-1 alpha Antibody maps to a region between residues 775 and the C-terminus (residue 826) of human hypoxia-inducible factor 1 [Uniprot# Q16665].
Reactivity Notes
Monkey (COS-7) and Rat reactivities were reported in customer review feedback. Detection of HIF1 alpha in both Mouse and Human tissue by IHC. Chicken reactivity was reported in scientific literature (PMID: 25632022). Canine reactivity reported in scientific literature (PMID: 28701694). Reactivity with Goat is reported in PMID: 21599540.
Clonality
Polyclonal
Host
Rabbit
Isotype
IgG
Theoretical MW
93 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Scientific Data Images for HIF-1 alpha Antibody
Simple Western: HIF-1 alpha Antibody [NB100-449]
Simple Western: HIF-1 alpha Antibody [NB100-449] - Simple Western lane view shows a specific band for HIF-1 alpha in 0.5 mg/ml of hypoxic HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.Applications for HIF-1 alpha Antibody
Application
Recommended Usage
Chromatin Immunoprecipitation (ChIP)
1:10-1:500
ELISA
1:100-1:2000
Flow Cytometry
0.125 ug per 1 million cells in a 150 mcl reaction
Immunocytochemistry/ Immunofluorescence
1:10-1:500
Immunohistochemistry
1:100 - 1:500
Immunohistochemistry-Frozen
1:50-1:200
Immunohistochemistry-Paraffin
1:50-1:200
Immunoprecipitation
2-5 ug/mg lysate
Simple Western
1:200
Western Blot
1:2000 - 1:10000
Application Notes
ChIP usage was reported in scientific literature (PMID: 25557133). In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: Tested in Hypoxic HeLa lysate 0.5 mg/mL, separated by Size, antibody dilution of 1:200, apparent MW was 115 kDa. For IHC-P, Tris-EDTA pH 9.0 buffer is recommended for the heat induced epitope retrieval. ELISA (PMID: 17556599 and 16966370). Knockout data (PMID: 31793879). Use in Flow-intracellular reported in scientific literature (PMID:31722203).
See Simple Western Antibody Database for Simple Western validation: Tested in Hypoxic HeLa lysate 0.5 mg/mL, separated by Size, antibody dilution of 1:200, apparent MW was 115 kDa. For IHC-P, Tris-EDTA pH 9.0 buffer is recommended for the heat induced epitope retrieval. ELISA (PMID: 17556599 and 16966370). Knockout data (PMID: 31793879). Use in Flow-intracellular reported in scientific literature (PMID:31722203).
Reviewed Applications
Read 29 reviews rated 4.3 using NB100-449 in the following applications:
Formulation, Preparation, and Storage
Purification
Immunogen affinity purified
Formulation
TBS, 0.1% BSA
Preservative
0.09% Sodium Azide
Concentration
0.2 mg/ml
Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store at 4C. Do not freeze.
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
Gene Symbol
HIF1A
UniProt
Additional HIF-1 alpha/HIF1A Products
Product Documents for HIF-1 alpha Antibody
Product Specific Notices for HIF-1 alpha Antibody
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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