HIF-1 alpha Antibody - BSA Free
Novus Biologicals, part of Bio-Techne | Catalog # NB100-654
Conjugate
Catalog #
Key Product Details
Validated by
Knockout/Knockdown, Orthogonal Validation, Biological Validation
Species Reactivity
Validated:
Human, Mouse, Rat, Porcine, Bovine
Cited:
Human, Mouse, Rat, Porcine, Bovine
Applications
Validated:
Gel Super Shift Assays, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Knockdown Validated, Simple Western, Western Blot
Cited:
IF/IHC, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Knockdown Validated, Western Blot
Label
Unconjugated
Antibody Source
Polyclonal Rabbit IgG
Format
BSA Free
Concentration
1.0 mg/ml
Product Specifications
Immunogen
This HIF-1 alpha Antibody was developed against a fusion protein containing amino acids 432 - 528 of human HIF-1 alpha [Uniprot# Q16665].
Reactivity Notes
Mouse reactivity was reported in scientific literature (PMID: 23959856). Rat reactivity was reported in scientific literature (PMID: 24122166). Use in Bovine reported in scientific literature (PMID:32054096).
Localization
Nuclear
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 - BSA Free
Western Blot: HIF-1 alpha AntibodyBSA Free [NB100-654]
Western Blot: HIF-1 alpha Antibody [NB100-654] - Effects of in vitro hypoxia on the expression of mouse HIF-1 alpha in HIF-1 alpha+/- and HIF-1 alpha+/+ astrocytes. Confluent astrocyte monolayers of both cell types were exposed to a 6 h in vitro hypoxia. HIF-1 alpha mRNA expression was determined by RT-PCR as described in Materials and Methods. Each bar represents the mean +/- SD of relative density/volumes of the bands on film negatives from at least three experiments. Asterisks and number sign indicate significant difference (p < 0.01; one-way ANOVA, followed by multiple comparisons among means). Image collected and cropped by CiteAb from the following publication (https://jneuroinflammation.biomedcentral.com/articles/10.1186/1742-2094-4-12), licensed under a CC-BY license.Western Blot: HIF-1 alpha AntibodyBSA Free [NB100-654]
Western Blot: HIF-1 alpha Antibody [NB100-654] - Effects of CoCl2 treatment on HIF-1 alpha expression in mouse astrocytes. Western blots using nuclear proteins show that both hypoxia (H) and CoCl2 (Co) up-regulated HIF-1 alpha protein in HIF-1 alpha+/+ and HIF-1 alpha+/- cells. There was no HIF-1 alpha protein detected in control cells. Image collected and cropped by CiteAb from the following publication (https://jneuroinflammation.biomedcentral.com/articles/10.1186/1742-2094-4-12), licensed under a CC-BY license.Immunohistochemistry-Paraffin: HIF-1 alpha Antibody - BSA Free [NB100-654]
Immunohistochemistry-Paraffin: HIF-1 alpha Antibody [NB100-654] - IHC analysis of a formalin-fixed paraffin-embedded tissue section of human endometrium carcinoma AN3CA cell line based xenograft using rabbit polyclonal HIF-1 alpha antibody NB100-654 at 1:300 dilution. The signal was developed using HRP-labelled secondary antibody and DAB reagent, and the section was further counterstained using hematoxylin. The tested section depicted mainly a diffused cytoplasmic staining but there were some cells which showed nuclear signal also (representing hypoxic cells).Applications for HIF-1 alpha Antibody - BSA Free
Application
Recommended Usage
Immunocytochemistry/ Immunofluorescence
1:10 - 1:500. Use reported in scientific literature
Immunohistochemistry
1:100 - 1:300
Immunohistochemistry-Frozen
1:100 - 1:300. Use reported in scientific literature
Immunohistochemistry-Paraffin
1:100 - 1:300. Use reported in scientific literature
Knockdown Validated
reported in scientific literature (PMID 32054096)
Simple Western
1:100
Western Blot
1:1000
Application Notes
Nuclear extracts are recommended.
In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
Please Note: Optimal dilutions of this antibody should be experimentally determined.
Reviewed Applications
Read 1 review rated 5 using NB100-654 in the following applications:
Formulation, Preparation, and Storage
Purification
Immunogen affinity purified
Formulation
PBS
Format
BSA Free
Preservative
0.02% Sodium Azide
Concentration
1.0 mg/ml
Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Aliquot and store at -20C or -80C. 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
Gene Symbol
HIF1A
Additional HIF-1 alpha/HIF1A Products
Product Documents for HIF-1 alpha Antibody - BSA Free
Product Specific Notices for HIF-1 alpha 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.
Loading...
Loading...
Loading...
Loading...