HIF-1 alpha (HIF1A) Rabbit Polyclonal Antibody

CAT#: TA336308

Rabbit Polyclonal HIF-1 alpha Antibody

Product Images

Staining of human placenta, villi.

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  Analysis of HIF-1 alpha in human kidney tumor.

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  Analysis in A. PC12 (rat) cells, 20% oxygen and B. PC12 (rat) cells, 1% oxygen.

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  Detection of HIF-1 Alpha (green) in RCC4 cells using TA336308. Nuclei (Blue) were counterstained using Hoechst 33258.

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  Analysis in a hypoxic sample. Lane 1: CoCl2 treated Cos-7 nuclear extract (50 ug, hypoxic). Lane 2: Untreated Cos-7 nuclear extract (50 ug, normoxic). 10 second ECL exposure.

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  Cofilin phosphorylation is HIF-1α dependent.Wt, shC, and the HIF-1α knock down cell clones c1 and c2 cells were lysed after 24 hrs of normoxia (20% O2) or hypoxia (1% O2). Cell extracts were analysed by Western blots. Note that p-cofilin levels are reduced in c2 and c2 cells compared to wt and shC cells at 20% O2 and 1% O2.

• Simple Western lane view shows lysates of MCF +/- DMOG, BioSpherix MSCs in hypoxic conditions, and BioSpherix MSCs in normoxic conditions loaded at 0.5 mg/ml. A band was detected for HIF-1 alpha at approximately 116 kDa (as indicated) using TA336308 (1:100 dilution) followed by Anti-Rabbit Secondary Antibody (042-206, ProteinSimple). This experiment was conducted under standard assay conditions, and using the 12-230 kDa separation module (SW-W004). Non-specific interaction with the 230 kDa Simple Western standard seen with this antibody. Image from an internal validation.

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Specifications

Product Data

• Applications: ChIP, FC, ICC/IF, IHC, Immunoblotting, IP, Simple Western, WB
• Recommended Dilution: Immunohistochemistry: 5 - 10 ug/mL, Immunohistochemistry-Frozen: 5 - 10 ug/mL, Immunohistochemistry-Paraffin: 5 - 10 ug/mL, Immunocytochemistry/ Immunofluorescence: 1:100, Immunoprecipitation: 1:10 - 1:500, Flow Cytometry: 1:10 - 1:1000, Immunoblotting, Immunohistochemistry Whole-Mount, Simple Western, Chromatin Immunoprecipitation (ChIP): 1:10-1:500, Western Blot: 1-2ug/ml
• Reactivities: Human, Mouse, Rat, Goat, Hamster, Rabbit, Primate
• Host: Rabbit
• Clonality: Polyclonal
• Immunogen: A fusion protein including residues 530-825 of the mouse HIF-1 alpha protein. [UniProt# Q61221]
• Formulation: Tris-citrate/phosphate, pH 7, 0.1% Sodium azide. Store at 4C. Do not freeze.
• Concentration: lot specific
• Purification: Immunogen affinity purified
• Conjugation: Unconjugated
• Storage: Store at -20°C as received.
• Stability: Stable for 12 months from date of receipt.
• Predicted Protein Size: 115 kDa
• Gene Name: hypoxia inducible factor 1 alpha subunit
• Database Link:
  NP_851397
  Entrez Gene 15251 MouseEntrez Gene 29560 RatEntrez Gene 3091 Human
  UniProt ID Q16665
• Background: HIF1 (hypoxia-inducible factor 1), a heterodimeric transcription factor complex central to cellular response to hypoxia, consists of two subunits (HIF-1 alpha and HIF-1 beta) which are basic helix-loop-helix proteins of the PAS (Per, ARNT, Sim) family. Expression of HIF-1 alpha protein is regulated by cellular oxygen level alterations as well as in oxygen-independent manner via different cytokines (through the PI3K-AKT-mTOR pathway), growth factors, oncogenic activation, or loss of tumor suppressor function etc. In normoxic cells, HIF-1 alpha is proline hydroxylated leading to a conformational change that promotes its binding to the VLH (von Hippel Lindau) protein E3 ligase complex; ubiquitination and followed by rapid proteasomal degradation. Hypoxia as well as chemical hydroxylase inhibitors (desferrioxamine, cobalt etc.) inhibit HIF-1 alpha degradation and lead to its accumulation in the cells, whereas, contrastingly, HIF-1 beta/ARNT (AhR nuclear translocator) remains stable under both conditions. Besides their critical role in hypoxic response, HIF1s regulates the transcription of genes responsible for angiogenesis, erythropoiesis/iron-metabolism, glucose metabolism, cell proliferation/survival, adipogenesis, carotid body formation, B lymphocyte development and immune reactions.
• Synonyms: bHLHe78; HIF-1-alpha; HIF-1A; HIF-1alpha; HIF1; HIF1-ALPHA; MOP1; PASD8
• Note: This HIF-1 alpha antibody is useful for Immunocytochemistry/Immunofluorescence, Immunohistochemistry on paraffin-embedded and frozen sections and Western blot, recognizing a band in hypoxic samples at ~115 kDa. Rat retina and brain cortex have been tested. Rat brain cortex is a bit dirty but this antibody does detect upregulation of HIF-1 alpha in hypoxic samples. HIF-1 alpha Protein NBC1-18422 has been successfully used for antibody preadsorption/blocking assay in IHC-P with Hif-1alpha antibody NB100-479 (refer to product review by Yves Heremans on the datasheet of NBC1-18422).

Reference Data

• Protein Families: Transcription Factors
• Protein Pathways: mTOR signaling pathway, Pathways in cancer, Renal cell carcinoma