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HIF-1 alpha Antibody - BSA Free

Novus Biologicals, part of Bio-Techne | Catalog # NB100-654

Novus Biologicals, part of Bio-Techne

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.

Scientific Data Images for HIF-1 alpha Antibody - BSA Free

Western Blot: HIF-1 alpha AntibodyBSA Free [NB100-654]

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 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 - 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.
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

Hypoxia contributes to the pathophysiology of human disease, including myocardial and cerebral ischemia, cancer, pulmonary hypertension, congenital heart disease and chronic obstructive pulmonary disease (1). In cancer and particularly solid tumors, hypoxia plays a critical role in the regulation of genes involved in stem cell renewal, epithelial to mesenchymal transition (EMT), metastasis and angiogenesis. In the tumor microenvironment (TME), hypoxia influences the properties and function of stromal cells (e.g., fibroblasts, endothelial and immune cells) and is a strong determinant of tumor progression (2,3).

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

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

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.

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