Hypoxia Inducible Factor 1, alpha Subunit (Basic Helix-Loop-Helix Transcription Factor) (HIF1A) (AA 432-528) antibody

Details for Product No. ABIN151066
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Antigen
Synonyms hif1a, AA959795, HIF1alpha, MOP1, bHLHe78, HIF-1A, HIF-1alpha, HIF1, HIF1-ALPHA, PASD8, HIF1-alpha, hif-1a
Epitope
AA 432-528
(41), (39), (23), (11), (9), (8), (8), (7), (7), (6), (6), (6), (6), (5), (2), (2), (2), (2), (2), (2), (2), (2), (2), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1)
Reactivity
Human
(260), (112), (76), (54), (36), (31), (21), (16), (16), (15), (10), (3), (2), (1), (1), (1)
Host
Rabbit
(188), (126), (2)
Clonality
Polyclonal
Conjugate
Un-conjugated
(27), (15), (12), (12), (12), (6), (3), (3), (2), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1)
Application
Western Blotting (WB), Chromatin Immunoprecipitation (ChIP), Gel Shift (GS), Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunoprecipitation (IP)
(235), (98), (91), (82), (78), (68), (46), (39), (13), (12), (10), (10), (9), (4), (3), (1), (1)
Pubmed 15 references available
Quantity 0.1 mL
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Catalog No. ABIN151066
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Immunogen Fusion protein made to an internal sequence of human HIF-1 alpha (containing amino acids 432-528)
Cross-Reactivity (Details) Canine reactivity reported in scientific literature (PMID: 24140705) Mouse results are mixed.
Purification affinity purified
Alternative Name HIF1A / HIF1 alpha
Background Hypoxia contributes significantly to the pathophysiology of major categories of humandisease, including myocardial and cerebral ischemia, cancer, pulmonary hypertension,congenital heart disease, and chronic obstructive pulmonary disease.HIF-1 is a nuclear protein involved in mammalian oxygen homeostasis. This occurs as aposttranslational modification by prolyl hydroxylation. HIF-1 is a heterodimer composedof HIF-1 alpha and HIF-1 beta subunits. Both subunits are constantly translated.However, under normoxic conditions, human HIF-1 alpha is hydroxylated at Pro402 orPro564 by a set of HIF prolyl hydroxylases, is polyubiquinated, and eventually degradedin proteosomes. Under hypoxic conditions, the lack of hydroxylation prevents HIFdegradation and increases transcriptional activity. Therefore, the concentration of HIF-1alpha increases in the cell. In contrast, HIF-1 beta remains stable under either condition.HIF hydroxylases provide insight into hypoxic cell responses, which may be used to helpisolate therapeutic targets. Alternate Names: anti-Hypoxia-inducible factor 1 alpha antibody, anti-HIF1 alpha antibody, anti-ARNTinteracting proteinantibody, anti-Hif1a antibody, anti-ARNT interacting protein antibody,anti-HIF-1alpha antibody, anti-Hypoxia inducible factor 1 alpha antibody, anti-Hypoxiainducible factor 1 alpha subunit basic helix antibody.
Gene Symbol: HIF1A
Gene ID 3091, 15251, 29560
UniProt Q16665
Application Notes This HIF-1 alpha antibody is useful for Chromatin Immunoprecipitation (PMID 22891341), Immunohistochemistry, Western Blot, Gel Shift Assays, Immunocytochemistry/Immunfluorescence (PMID: 21270095, 21270095), and Immunoprecipitation. In Western blot a band is observed ~120 kDa. IHC applications have been mixed on mouse. For ChIP, nuclear extracts are recommended. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended.
Recommended dilutions: Chromatin Immunoprecipitation 25 µL for 15 million cells, Gel Super Shift Assays 1:1-1:100, Immunocytochemistry/Immunofluorescence 1:10-1:500, Immunohistochemistry 1:10-1:500, Immunohistochemistry-Frozen 1:10-1:500, Immunohistochemistry-Paraffin 1:100, Immunoprecipitation 1:1000, Western Blot 1:500-1:1000
Protocol Western blot Protocol specific for HIF-1 alpha antibody Western Blot Protocol
1. Perform SDS-PAGE on hypoxic samples to be analyzed, loading 30 µg of nuclear protein per lane.
. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
. Rinse the blot.
. Block the membrane using standard blocking buffer for at least 1 hour.
. Wash the membrane in wash buffer three times for 10 minutes each.
. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature.
. Wash the membrane in wash buffer three times for 10 minutes each.
. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
. Apply the detection reagent of choice in accordance with the manufacturers instructions.Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2 %.Immunohistochemistry Protocol for HIF-1 alpha antibody Immunohistochemistry-Paraffin Embedded SectionsAntigen Unmasking:Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.
0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes. Staining:
. Wash sections in deionized water three times for 5 minutes each.
. Wash sections in wash buffer for 5 minutes.
. Block each section with 100-400 µL blocking solution for 1 hour at room temperature.
. Remove blocking solution and add 100-400 µL diluted primary antibody. Incubate overnight at 4 C.
. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
. Add 100-400 µL biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
. Add 100-400 µL Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
. Wash sections three times in wash buffer for 5 minutes each.
. Add 100-400 µL DAB substrate to each section and monitor staining closely.
. As soon as the sections develop, immerse slides in deionized water.
. Counterstain sections in hematoxylin.
. Wash sections in deionized water two times for 5 minutes each.
. Dehydrate sections.
. Mount coverslips.
Restrictions For Research Use only
Format Liquid
Concentration 1 mg/mL
Buffer Tris-glycine, 150 mM NaCl, Sodium Azide
Preservative Sodium azide
Precaution of Use WARNING: Reagents contain sodium azide. Sodium azide is very toxic if ingested or inhaled. Avoid contact with skin, eyes, or clothing. Wear eye or face protection when handling. If skin or eye contact occurs, wash with copious amounts of water. If ingested or inhaled, contact a physician immediately. Sodium azide yields toxic hydrazoic acid under acidic conditions. Dilute azide-containing compounds in running water before discarding to avoid accumulation of potentially explosive deposits in lead or copper plumbing.
Handling Advice Avoid freeze-thaw cycles
Storage 4 °C
Storage Comment 4 °C short term. Aliquot and store at -20 °C long term.
Supplier Images
anti-Hypoxia Inducible Factor 1, alpha Subunit (Basic Helix-Loop-Helix Transcription Factor) (HIF1A) (AA 432-528) antibody Western Blot analysis of ABIN151066, anti-HIF-1 alpha, on normoxic and hypoxic nuclear rat cell lysates.
anti-Hypoxia Inducible Factor 1, alpha Subunit (Basic Helix-Loop-Helix Transcription Factor) (HIF1A) (AA 432-528) antibody (2) Staining of human kidney, renal tubular epithelium in Cortex using ABIN151066.
General Bergeron, Gidday, Yu et al.: "Role of hypoxia-inducible factor-1 in hypoxia-induced ischemic tolerance in neonatal rat brain." in: Annals of neurology, Vol. 48, Issue 3, pp. 285-96, 2000 (PubMed).

Liu, Wu, Cai et al.: "Hypoxic preconditioning of cardiomyocytes and cardioprotection: phophorylation of HIF-1alpha induced by p42/p44 mitogen-activated protein kinases is involved." in: Pathophysiology : the official journal of the International Society for Pathophysiology / ISP, Vol. 9, Issue 4, pp. 201-205, 2003 (PubMed).

Ietta, Wu, Winter et al.: "Dynamic HIF1A regulation during human placental development." in: Biology of reproduction, Vol. 75, Issue 1, pp. 112-21, 2006 (PubMed).

Harris, Brown, Jorgensen et al.: "Bone marrow-derived cells home to and regenerate retinal pigment epithelium after injury." in: Investigative ophthalmology & visual science, Vol. 47, Issue 5, pp. 2108-13, 2006 (PubMed).

Wang, Biju, Wang et al.: "Cytoprotective effects of hypoxia against cisplatin-induced tubular cell apoptosis: involvement of mitochondrial inhibition and p53 suppression." in: Journal of the American Society of Nephrology : JASN, Vol. 17, Issue 7, pp. 1875-85, 2006 (PubMed).

Lin, Lee, Yun: "Differentiation arrest by hypoxia." in: The Journal of biological chemistry, Vol. 281, Issue 41, pp. 30678-83, 2006 (PubMed).

Liao, Corle, Seagroves et al.: "Hypoxia-inducible factor-1alpha is a key regulator of metastasis in a transgenic model of cancer initiation and progression." in: Cancer research, Vol. 67, Issue 2, pp. 563-72, 2007 (PubMed).

Harvey, Kind, Thompson: "Regulation of gene expression in bovine blastocysts in response to oxygen and the iron chelator desferrioxamine." in: Biology of reproduction, Vol. 77, Issue 1, pp. 93-101, 2007 (PubMed).

Young, Williams, Chandra et al.: "Central role of Muc5ac expression in mucous metaplasia and its regulation by conserved 5' elements." in: American journal of respiratory cell and molecular biology, Vol. 37, Issue 3, pp. 273-90, 2007 (PubMed).

Kim, Gao, Liu et al.: "Hypoxia-inducible factor 1 and dysregulated c-Myc cooperatively induce vascular endothelial growth factor and metabolic switches hexokinase 2 and pyruvate dehydrogenase kinase 1." in: Molecular and cellular biology, Vol. 27, Issue 21, pp. 7381-93, 2007 (PubMed).

Ohtomo, Nangaku, Izuhara et al.: "Cobalt ameliorates renal injury in an obese, hypertensive type 2 diabetes rat model." in: Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association, Vol. 23, Issue 4, pp. 1166-72, 2008 (PubMed).

Sikder, Kodadek: "The neurohormone orexin stimulates hypoxia-inducible factor-1 activity." in: Genes & development, Vol. 21, Issue 22, pp. 2995-3005, 2007 (PubMed).

Piantadosi, Suliman: "Transcriptional Regulation of SDHa flavoprotein by nuclear respiratory factor-1 prevents pseudo-hypoxia in aerobic cardiac cells." in: The Journal of biological chemistry, Vol. 283, Issue 16, pp. 10967-77, 2008 (PubMed).

Singh, Gu, Agarwal: "Silibinin inhibits colorectal cancer growth by inhibiting tumor cell proliferation and angiogenesis." in: Cancer research, Vol. 68, Issue 6, pp. 2043-50, 2008 (PubMed).

Lauzier, Robitaille, Chan et al.: "(2R)-[(4-Biphenylylsulfonyl)amino]-N-hydroxy-3-phenylpropionamide (BiPS), a matrix metalloprotease inhibitor, is a novel and potent activator of hypoxia-inducible factors." in: Molecular pharmacology, Vol. 74, Issue 1, pp. 282-8, 2008 (PubMed).

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