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HIF-1 alpha Antibody (H1alpha67)

Novus Biologicals, part of Bio-Techne | Catalog # NB100-123

Novus Biologicals, part of Bio-Techne

Key Product Details

Validated by

Knockout/Knockdown, Orthogonal Validation, Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat, Porcine, Avian, Bovine, Canine, Ferret, Primate, Rabbit, Sheep

Cited:

Human, Mouse, Rat, Porcine, Ovine, Primate, Rabbit

Applications

Validated:

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

Cited:

Chemotaxis, ELISA, Flow Cytometry, Gel Supershift Assay, IF/IHC, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunoprecipitation, Knockout Validated, Simple Western, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # H1alpha67

Concentration

1.0 mg/ml

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.

Scientific Data Images for HIF-1 alpha Antibody (H1alpha67)

Immunohistochemistry: HIF-1 alpha Antibody (H1alpha67) [NB100-123]

Immunohistochemistry: HIF-1 alpha Antibody (H1alpha67) [NB100-123]

Immunohistochemistry: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - Results of in situ hybridization and immunohistochemistry on thin adjacent section to detect expression of HIF-1a1.2 mRNA and HIF1a protein in malignant and benign prostate tissue. In situ hybridization (antisense probe, Fig. 3a) and immunostaining with HIF1a Ab2 (Fig. 3c) on thin adjacent sections of NE-differentiated prostate adenocarcinoma showed co-localization of HIF1a1.2 transcript and HIF-1a protein. Incubation with sense probe did not generate any detectable hybridization signals (Fig. 3b). Both In situ hybridization (Fig. 3d antisense) and HIF-1a Ab2 immunostaining (Fig. 3f) were negative in non-NE-differentiated prostate adenocarcinoma. In situ hybridization with sense probe performed on non-NE-differentiated prostate cancer (Fig. 3e). Image collected and cropped by CiteAb from the following publication (https://www.biomedcentral.com/1471-2407/10/385), licensed under a CC-BY license.
Knockdown Validated: HIF-1 alpha Antibody (H1alpha67) [NB100-123]

Knockdown Validated: HIF-1 alpha Antibody (H1alpha67) [NB100-123]

Knockdown Validated: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - HIF-1 inhibits the apoptosis of hypoxic glioblastoma cells. U87MG cells were transfected with siRNA against HIF-1alpha. Forty-eight hours after transfection, cells were incubated for 2 h in hypoxia (1% O2). HIF-1alpha was detected by immunoblotting. beta-Actin was used as a loading control. The results are representative for three independent experiments. Image collected and cropped by CiteAb from the following publication (https://www.nature.com/articles/cddis2013562) licensed under a CC-BY license.
Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-123]

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-123]

Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-123] - MLN4924 induces accumulation of HIF1a in a time dependent manner. Cells were treated with 0.1uM MLN4924 for indicated time periods, followed by IB with indicated antibodies. Image collected and cropped by CiteAb from the following publication (https://www.nature.com/articles/cddis2012125) licensed under a CC-BY license.

Applications for HIF-1 alpha Antibody (H1alpha67)

Application
Recommended Usage

Chromatin Immunoprecipitation

1:10 - 1:500. Use reported in scientific literature

Chromatin Immunoprecipitation (ChIP)

1:10-1:500

ELISA

1:100 - 1:2000. Use reported in scientific literature

Flow Cytometry

1:10 - 1:1000

Gel Super Shift Assays

1:1 - 1:100. Use reported in scientific literature

Immunoblotting

reported in multiple pieces of scientific literature

Immunohistochemistry

1:100 - 1:300

Immunohistochemistry-Paraffin

1:100 - 1:300

Immunoprecipitation

1:10

Knockout Validated

reported in scientific literature (PMID 27991597)

Western Blot

1:500 - 1:1000
Application Notes
By WB, this antibody recognizes bands at 120kDa representing HIF-1 alpha in induced tissues and cells. Multiple bands may be seen at 120kDa representing post-translational modifications. Nuclear extracts are recommended for WB.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 10 reviews rated 4.2 using NB100-123 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS with 1% BSA

Preservative

0.05% 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

Store at 4C short term. Aliquot and store at -20C long term. 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, H1alpha67

Gene Symbol

HIF1A

Additional HIF-1 alpha/HIF1A Products

Product Documents for HIF-1 alpha Antibody (H1alpha67)

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 (H1alpha67)

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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