HIF-1 alpha Antibody (2443B) - BSA Free

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-75977

Recombinant Monoclonal Antibody.
Novus Biologicals, part of Bio-Techne
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NBP2-75977
NBP2-75977SS
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Key Product Details

Validated by

Knockout/Knockdown, Biological Validation

Species Reactivity

Human

Applications

Immunocytochemistry/ Immunofluorescence, Knockout Validated, Simple Western, Western Blot

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # 2443B

Format

BSA Free

Concentration

1.0 mg/ml

View all HIF-1 alpha/HIF1A Antibodies »

Product Specifications

Immunogen

This recombinant HIF-1 alpha Antibody (2443B) was developed against partial recombinant human HIF-1 alpha protein (amino acids 1-826) [Uniprot# Q16665].

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for HIF-1 alpha Antibody (2443B) - BSA Free

Western Blot: HIF-1 alpha Antibody (2443B)BSA Free [NBP2-75977]

Western Blot: HIF-1 alpha Antibody (2443B)BSA Free [NBP2-75977]

Western Blot: HIF-1 alpha Antibody (2443B) [NBP2-75977] - Total protein from Human U87 cells treated with or without CoCl2 and under Normoxic and Hypoxic conditions was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/ml anti-HIF-1 alpha in 5% non-fat milk in TBST and detected with an anti-rabbit HRP secondary antibody using chemiluminescence.
Knockout Validated: HIF-1 alpha Antibody (2443B) - BSA Free [NBP2-75977]

Knockout Validated: HIF-1 alpha Antibody (2443B) - BSA Free [NBP2-75977]

Knockout Validated: HIF-1 alpha Antibody (2443B) [NBP2-75977] - Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and HIF-1 alpha knockout HeLa cell line (KO) untreated (-) or treated (+) with 1 mM DFO for overnight. PVDF membrane was probed with 2 ug/mL of Rabbit Anti-Human HIF-1 alpha Monoclonal Antibody (NBP2-75977) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for HIF-1 alpha at an approximatel molecular weight of 120 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # MAB5718) is shown as a loading control.This experiment was conducted under reducing conditions.
Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody (2443B) - BSA Free [NBP2-75977]

Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody (2443B) - BSA Free [NBP2-75977]

Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody (2443B) [NBP2-75977] - Immunofluorescent staining of HIF-1 alpha using catalog number NBP2-75977 (clone 2443B). Staining was observed in Hela cells treated with DFO but not in untreated Hela cells.
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Applications for HIF-1 alpha Antibody (2443B) - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

Optimal dilutions of this antibody should be experimentally determined.

Knockout Validated

Optimal dilutions of this antibody should be experimentally determined.

Simple Western

Optimal dilutions of this antibody should be experimentally determined.

Western Blot

Optimal dilutions of this antibody should be experimentally determined.
Application Notes
Simple western reported by internal validaton.

Formulation, Preparation, and Storage

Purification

Protein A or G 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

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, anti-HIF-1 alpha, anti-HIF1A, H1alpha67

Gene Symbol

HIF1A

Additional HIF-1 alpha/HIF1A Products

Product Documents for HIF-1 alpha Antibody (2443B) - BSA Free

Product Datasheets

Certificate of Analysis

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Product Specific Notices for HIF-1 alpha Antibody (2443B) - 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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