HIF-1 alpha Antibody (H1alpha67) [Biotin]
Novus Biologicals, part of Bio-Techne | Catalog # NB100-123B
Conjugate
Catalog #
Key Product Details
Validated by
Biological Validation
Species Reactivity
Human, Mouse, Rat, Porcine, Avian, Bovine, Canine, Ferret, Primate, Rabbit, Sheep
Applications
Chromatin Immunoprecipitation, Chromatin Immunoprecipitation (ChIP), ELISA, Flow Cytometry, Gel Super Shift Assays, Immunoblotting, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunoprecipitation, Knockdown Validated, Knockout Validated, Western Blot
Label
Biotin
Antibody Source
Monoclonal Mouse IgG2B Clone # H1alpha67
Concentration
Please see the vial label for concentration. If unlisted please contact technical services.
Product Specifications
Immunogen
This HIF-1 alpha Antibody (H1alpha67) was developed against a fusion protein containing amino acids 432 - 528 of human HIF-1 alpha [Uniprot# Q16665].
Reactivity Notes
Please note that this antibody is reactive to Mouse and derived from the same host, Mouse. Additional Mouse on Mouse blocking steps may be required for IHC and ICC experiments. Please contact Technical Support for more information. Rabbit reactivity reported in scientific literature (PMID: 16738327, 26339038).
Localization
HIF-1 alpha is a nuclear protein that activates gene transcription in response to reduced cellular O2 concentration.
Clonality
Monoclonal
Host
Mouse
Isotype
IgG2B
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 (H1alpha67) [Biotin]
Western Blot: HIF-1 alpha Antibody (H1alpha67) [Biotin] [NB100-123B] - On day 1, MEF cells (+/+,-/-), were grown on 15cm dish (2x10 to the 6th cells). On day 2, cells were exposed to hypoxia for 4hrs. Cells were washed with ice cold PBS twice and whole cell protein was extracted with RIPA buffer fortified with protease. Upon quantification, 100ug of protein was fractionated on 7% polyacralymide gel. Gel was transferred overnight onto nitrocellulose membrane. The membrane was probed with HIF-1 alpha monoclonal antibody at a 1:500 dilution (NB100-123). The secondary antibody was conjugated with HRP and was used at a 1:2500 dilution. Photo courtesy of Dr. Gregg Semenza, Johns Hopkins University.
Applications for HIF-1 alpha Antibody (H1alpha67) [Biotin]
Application
Recommended Usage
Chromatin Immunoprecipitation
Optimal dilutions of this antibody should be experimentally determined.
Chromatin Immunoprecipitation (ChIP)
Optimal dilutions of this antibody should be experimentally determined.
ELISA
Optimal dilutions of this antibody should be experimentally determined.
Flow Cytometry
Optimal dilutions of this antibody should be experimentally determined.
Gel Super Shift Assays
Optimal dilutions of this antibody should be experimentally determined.
Immunoblotting
Optimal dilutions of this antibody should be experimentally determined.
Immunocytochemistry/ Immunofluorescence
Optimal dilutions of this antibody should be experimentally determined.
Immunohistochemistry
Optimal dilutions of this antibody should be experimentally determined.
Immunohistochemistry-Paraffin
Optimal dilutions of this antibody should be experimentally determined.
Immunoprecipitation
Optimal dilutions of this antibody should be experimentally determined.
Knockdown Validated
Optimal dilutions of this antibody should be experimentally determined.
Knockout Validated
Optimal dilutions of this antibody should be experimentally determined.
Western Blot
Optimal dilutions of this antibody should be experimentally determined.
Application Notes
Optimal dilution of this antibody should be experimentally determined.
Formulation, Preparation, and Storage
Purification
Protein A purified
Formulation
PBS
Preservative
0.05% Sodium Azide
Concentration
Please see the vial label for concentration. If unlisted please contact technical services.
Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store at 4C in the dark.
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, H1alpha67
Gene Symbol
HIF1A
Additional HIF-1 alpha/HIF1A Products
Product Documents for HIF-1 alpha Antibody (H1alpha67) [Biotin]
Product Specific Notices for HIF-1 alpha Antibody (H1alpha67) [Biotin]
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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