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

Details for Product No. ABIN151033, Supplier: Log in to see
Antigen
  • hif1a
  • AA959795
  • HIF1alpha
  • MOP1
  • bHLHe78
  • HIF-1A
  • HIF-1alpha
  • HIF1
  • HIF1-ALPHA
  • PASD8
  • HIF1-alpha
  • hif-1a
  • hypoxia inducible factor 1 alpha
  • hypoxia inducible factor 1, alpha subunit
  • hypoxia inducible factor 1, alpha subunit (basic helix-loop-helix transcription factor)
  • hypoxia-inducible factor 1 alpha
  • hypoxia-inducible factor 1, alpha subunit (basic helix-loop-helix transcription factor)
  • Protein HIF-1
  • hif-1a
  • Hif1a
  • HIF1A
  • hif-1
Alternatives
anti-Human Hypoxia Inducible Factor 1, alpha Subunit (Basic Helix-Loop-Helix Transcription Factor) antibody for Electrophoretic Mobility-Shift Assay
Reactivity
Cow (Bovine), Ferret, Human, Monkey, Mouse (Murine), Pig (Porcine), Primate, Rabbit, Rat (Rattus), Sheep (Ovine), Xenopus laevis, Yeast
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1
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1
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1
Host
Mouse
384
279
2
Clonality (Clone)
Monoclonal ()
Conjugate
Un-conjugated
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33
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25
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12
6
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1
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1
1
1
Application
Chromatin Immunoprecipitation (ChIP), ELISA, Flow Cytometry (FACS), Gel Shift (GS), Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunoprecipitation (IP), Ligand Activation (Act (ligand)), Substrate (S), Western Blotting (WB)
519
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107
91
62
16
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10
7
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3
3
2
2
2
1
1
Options
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Immunogen A fusion protein containing amino acids 432 - 528 of human HIF-1 alpha [UniProt# Q16665].
Clone H1alpha67
Isotype IgG2b
Purification Protein G purified
Alternative Name HIF-1 alpha (HIF1A Antibody Abstract)
Background Gene Symbol: HIF1A
Molecular Weight Theoretical MW: 93 kDa
Gene ID 3091
UniProt Q16665
Pathways Positive Regulation of Peptide Hormone Secretion, Regulation of Hormone Metabolic Process, Regulation of Hormone Biosynthetic Process, Cellular Response to Molecule of Bacterial Origin, Carbohydrate Homeostasis, Transition Metal Ion Homeostasis
Application Notes Western Blot 1:500, Chromatin Immunoprecipitation 1 - 5 μg/IP, Flow Cytometry 1:10 - 1:1000, ELISA 1:100 - 1:2000, Immunohistochemistry 1:20 - 1:50, Immunocytochemistry/Immunofluorescence 1:50, Immunoprecipitation 1:10 - 1:500, Immunohistochemistry-Paraffin 1:20 - 1:50, Immunohistochemistry-Frozen 1:20 - 1:50, Gel Super Shift Assays 1:1 - 1:100, Tissue Culture Substratum, Ligand ActivationChIP usage was reported in scientific literature. ELISA usage was reported in scientific literature (PMID: 20042684). Gel Super Shift Assays usage was reported in scientif literature (PMID: 22411794). Ligand Activation usage was reported in scientific literature (PMID: 26147748). IHC-P: No detection of Mouse is reported. WB: Detection of Mouse protein. In WB, a band can be seen at 120 kDa representing HIF-1 alpha in induced tissues and cells. Multiple bands may be seen at 100-120 kDa representing post-translational modification of HIF-1 alpha. For WB, testing on nuclear extracts is recommended. 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.
Comment

The antibodies are intended for use in vitro experiments only. Our antibodies have not been tested nor are recommended for use in vivo.

Protocol Western Blot Protocols specific for HIF-1 alpha Antibody HIF-1 alpha Western Blot General Information:
1. The HIF proteins are among the most rapidly degrading proteins ever studied. Upon cellular re-oxygenation it can be completely degraded in less than 1 minute. Therefore, it is critical to prep only a few plates/dishes/flasks of cells at a time and to immediately place the cells into ice cold buffers and perform the whole protein prep on ice.
. HIF-1 is largely undetectable in cells or tissues grown under normoxic conditions. It is stabilized only at O2 concentrations below 5 % or with treatment using certain agents (CoCl2, DFO, etc.) so proper sample preparation is critical.
. Upon stabilization HIF-1 translocates to the nucleus. The best western blots (cleanest) are always done using nuclear extracts. It is possible to detect HIF-1 in whole cell extracts, but they tend to be much dirtier and the staining is much weaker.
. Finally, we recommend that a positive/negative control always be run side by side so that it is possible to discern which band is upregulated in the hypoxic sample. Unprocessed HIF1 is ~95 kDa while the fully post-translationally modified form is ~116 kDa, or larger. Additionally, HIF-1 alpha may form a heterodimer with HIF-1 beta (Duan, et al. Circulation. 2005,111:2227-2232.).Depending on the sample, treatment, etc. you may see either a band or a doublet."EPO transcription can be activated by exposure of Hep3B cells to either hypoxia or cobalt chloride (7). HIF-1 binding activity was induced after 1 h and was maximal after 4-h treatment of Hep3B cells with 75 ,M cobalt chloride (Fig. 2A), which is similar to the kinetics of HIF-1 induction by hypoxia (data not shown). Exposure of HeLa cells to cobalt chloride for 4 h also induced HIF-1 activity. In contrast to hypoxia, which induced a doublet band corresponding to HIF-1 in EMSAs, cobalt chloride induced a single band of HIF-1 activity in both Hep3B and HeLa cells (compare Figs. 1A and 2A). We have not determined the basis for this reproducible difference in response to stimulation by hypoxia as compared to cobalt chloride" (Wang G, et al. (1993) PNAS 90, 4304-4308.).Thus, it is critical to be able to look at upregulation compared to the control.Western Blot Protocol 1 (used to produce the image on the datasheet)
. Perform SDS-PAGE (3-8 %) on samples to be analyzed, loading 40ug of total protein per lane (COS-7 treated and untreated lysates.
. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer apparatus.
. Stain the blot using ponceau S for 1-2 minutes to access the transfer of proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations and locations of molecular weight markers using a pencil.
. Rinse the blot in TBS for approximately 5 minutes.
. Block the membrane using 5 % non-fat dry milk in TBS for 1 hour.
. Dilute the mouse anti-HIF-1 alpha primary antibody in blocking buffer and incubate 2 hours at room temperature.
. Wash the membrane in water for 5 minutes and apply the diluted mouse-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) and incubate 1 hour at room temperature.
. Wash the blot in TBS containing 0.05-0.1 % Tween-20 for 10-20 minutes.
. Wash the blot in type I water for an additional 10-20 minutes (this step can be repeated as required to reduce background.
. Apply the detection reagent of choice in accordance with the manufacturers instructions (Amersham ECL is the standard reagent used).Note: Tween-20 can be added to the blocking buffer at a final concentration of 0.05-0.2 %, provided it does not interfere with antibody-antigen binding. Western Blot Procedure 2
Restrictions For Research Use only
Format Liquid
Concentration 1.0 mg/mL
Buffer PBS with 1 % BSA
Buffer contains: 0.05 % Sodium Azide
Preservative Sodium azide
Precaution of Use This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice Do not freeze.
Storage 4 °C,-20 °C
Storage Comment Store at 4°C short term. Aliquot and store at -20°C long term. Avoid freeze-thaw cycles.
Supplier Images
Immunohistochemistry (IHC) image for anti-Hypoxia Inducible Factor 1, alpha Subunit (Basic Helix-Loop-Helix Transcription Factor) (HIF1A) antibody (ABIN151033) Immunohistochemistry staining of human glioblastoma multiforme
Western Blotting (WB) image for anti-Hypoxia Inducible Factor 1, alpha Subunit (Basic Helix-Loop-Helix Transcription Factor) (HIF1A) antibody (ABIN151033) Western Blot analysis of 50ug cobalt chloride induced COS-7 nuclear extracts (NB800-P...
Western Blotting (WB) image for anti-Hypoxia Inducible Factor 1, alpha Subunit (Basic Helix-Loop-Helix Transcription Factor) (HIF1A) antibody (ABIN151033) Western Blot: HIF-1 alpha Antibody (H1alpha67) [ABIN151033] - HIF-1 alpha induction b...
Flow Cytometry (FACS) image for anti-Hypoxia Inducible Factor 1, alpha Subunit (Basic Helix-Loop-Helix Transcription Factor) (HIF1A) antibody (ABIN151033) Flow Cytometry: HIF-1 alpha Antibody (H1alpha67) [ABIN151033] - Analysis using the Al...
Flow Cytometry (FACS) image for anti-Hypoxia Inducible Factor 1, alpha Subunit (Basic Helix-Loop-Helix Transcription Factor) (HIF1A) antibody (ABIN151033) Flow Cytometry: HIF-1 alpha Antibody (H1alpha67) [ABIN151033] - Analysis using the Al...
Western Blotting (WB) image for anti-Hypoxia Inducible Factor 1, alpha Subunit (Basic Helix-Loop-Helix Transcription Factor) (HIF1A) antibody (ABIN151033) Western Blot: HIF-1 alpha Antibody (H1alpha67) [ABIN151033] - Analysis of HIF-1 alpha...
Immunohistochemistry (IHC) image for anti-Hypoxia Inducible Factor 1, alpha Subunit (Basic Helix-Loop-Helix Transcription Factor) (HIF1A) antibody (ABIN151033) Immunohistochemistry: HIF-1 alpha Antibody (H1alpha67) [ABIN151033] - Staining of HIF...
Immunofluorescence (IF) image for anti-Hypoxia Inducible Factor 1, alpha Subunit (Basic Helix-Loop-Helix Transcription Factor) (HIF1A) antibody (ABIN151033) Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody (H1alpha67) [ABIN151033]...
Product cited in: Hung, Chang, Huang, Yin, Hwang, Yang, Yang: "Inhibitor of Differentiation-1 and Hypoxia-Inducible Factor-1 Mediate Sonic Hedgehog Induction by Amyloid Beta-Peptide in Rat Cortical Neurons." in: Molecular neurobiology, Vol. 53, Issue 2, pp. 793-809, 2016 (PubMed). (Sample species: Rat (Rattus)). Further details: Chromatin Immunoprecipitation

Zhang, Takara, Yamakawa, Kidoya, Takakura: "Apelin as a marker for monitoring the tumor vessel normalization window during antiangiogenic therapy." in: Cancer science, Vol. 107, Issue 1, pp. 36-44, 2016 (PubMed). (Sample species: Mouse (Murine)). Further details: Western Blotting

Aiken, Roudier, Ciccone, Drouin, Stromberg, Vojnovic, Olfert, Haas, Gustafsson, Grenier, Birot: "Phosphorylation of murine double minute-2 on Ser166 is downstream of VEGF-A in exercised skeletal muscle and regulates primary endothelial cell migration and FoxO gene expression." in: FASEB journal : official publication of the Federation of American Societies for Experimental Biology, Vol. 30, Issue 3, pp. 1120-34, 2016 (PubMed). Further details: Western Blotting

Lee, Frattini, Bansal, Castano, Sherman, Hutchinson, Bruce, Califano, Liu, Cardozo, Iavarone, Lasorella: "An ID2-dependent mechanism for VHL inactivation in cancer." in: Nature, Vol. 529, Issue 7585, pp. 172-7, 2016 (PubMed). (Sample species: Human). Further details: Western Blotting

Maugeri, DAmico, Reitano, Saccone, Federico, Cavallaro, DAgata: "Parkin modulates expression of HIF-1α and HIF-3α during hypoxia in gliobastoma-derived cell lines in vitro." in: Cell and tissue research, Vol. 364, Issue 3, pp. 465-74, 2016 (PubMed). (Sample species: Human). Further details: Immunocytochemistry,Western Blotting,Immunofluorescence

Chen, Zeng, Xiao, Huang, Hu, Yao, Yu, Xiao, Xu, Ye: "Regulation of glucose metabolism by p62/SQSTM1 through HIF1α." in: Journal of cell science, Vol. 129, Issue 4, pp. 817-30, 2016 (PubMed). (Sample species: Human). Further details: Western Blotting

Ai, Chen, Li, Yang, Xu, Gui, Liu, Bai, Chen: "Danshen improves damaged cardiac angiogenesis and cardiac function induced by myocardial infarction by modulating HIF1α/VEGFA signaling pathway." in: International journal of clinical and experimental medicine, Vol. 8, Issue 10, pp. 18311-8, 2016 (PubMed).

Mazar, Qi, Lee, Marchica, Govindarajan, Shelley, Li, Ray, Perera: "MicroRNA 211 Functions as a Metabolic Switch in Human Melanoma Cells." in: Molecular and cellular biology, Vol. 36, Issue 7, pp. 1090-108, 2016 (PubMed). (Sample species: Human). Further details: Western Blotting

Mori, Yao, Learman, Kurozumi, Ishida, Ramakrishnan, Overmyer, Xue, Cawthorn, Reid, Taylor, Ning, Shah, MacDougald: "Induction of WNT11 by hypoxia and hypoxia-inducible factor-1α regulates cell proliferation, migration and invasion." in: Scientific reports, Vol. 6, pp. 21520, 2016 (PubMed). (Sample species: Human). Further details: Western Blotting

Luo, Wang, Hou, Hu, Jia, Qin, Zuo, Liu, Luo, Song, Wang, Pang: "ATP-Dependent Lon Protease Contributes to Helicobacter pylori-Induced Gastric Carcinogenesis." in: Neoplasia (New York, N.Y.), Vol. 18, Issue 4, pp. 242-52, 2016 (PubMed). (Sample species: Human). Further details: Western Blotting

Guinot, Lehmann, Wild, Frew: "Combined deletion of Vhl, Trp53 and Kif3a causes cystic and neoplastic renal lesions." in: The Journal of pathology, Vol. 239, Issue 3, pp. 365-73, 2016 (PubMed). (Sample species: Mouse (Murine)). Further details: Immunocytochemistry,Immunofluorescence,Immunohistochemistry

Bellanti, Mirabella, Mitarotonda, Blonda, Tamborra, Cinnella, Fersini, Ambrosi, Dambrosio, Vendemiale, Serviddio: "Propofol but not sevoflurane prevents mitochondrial dysfunction and oxidative stress by limiting HIF-1α activation in hepatic ischemia/reperfusion injury." in: Free radical biology & medicine, Vol. 96, pp. 323-33, 2016 (PubMed).

Badar, Handisides, Benito, Richie, Borthakur, Jabbour, Harutyunyan, Konoplev, Faderl, Kroll, Andreeff, Pearce, Kantarjian, Cortes, Thomas, Konopleva: "Phase I study of evofosfamide, an investigational hypoxia-activated prodrug, in patients with advanced leukemia." in: American journal of hematology, Vol. 91, Issue 8, pp. 800-5, 2016 (PubMed). (Sample species: Human). Further details: Immunohistochemistry (Paraffin-embedded Sections)

Fecher, Horwath, Friedrich, Rupp, Deepe: "Inverse Correlation between IL-10 and HIF-1α in Macrophages Infected with Histoplasma capsulatum." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 197, Issue 2, pp. 565-79, 2016 (PubMed). (Sample species: Mouse (Murine)). Further details: Western Blotting

Lo Dico, Costa, Martelli, Diceglie, Rajata, Rizzo, Mancone, Tripodi, Ottobrini, Alessandro, Conigliaro: "MiR675-5p Acts on HIF-1α to Sustain Hypoxic Responses: A New Therapeutic Strategy for Glioma." in: Theranostics, Vol. 6, Issue 8, pp. 1105-18, 2016 (PubMed). Further details: Immunocytochemistry,Immunohistochemistry (Paraffin-embedded Sections),Immunofluorescence

Samanta, Park, Andrabi, Shelton, Gilkes, Semenza: "PHGDH Expression Is Required for Mitochondrial Redox Homeostasis, Breast Cancer Stem Cell Maintenance, and Lung Metastasis." in: Cancer research, Vol. 76, Issue 15, pp. 4430-42, 2016 (PubMed). (Sample species: Human). Further details: Western Blotting

Hoefflin, Lahrmann, Warsow, Hübschmann, Spath, Walter, Chen, Hofer, Macher-Goeppinger, Tolstov, Korzeniewski, Duensing, Grüllich, Jäger, Perner, Schönberg, Nyarangi-Dix, Isaac, Hatiboglu, Teber et al.: "Spatial niche formation but not malignant progression is a driving force for intratumoural heterogeneity. ..." in: Nature communications, Vol. 7, pp. ncomms11845, 2016 (PubMed). Further details: Immunohistochemistry

Sanagawa, Iwaki, Asai, Sakakibara, Norimoto, Sobel, Fujii: "Sphingosine 1‑phosphate induced by hypoxia increases the expression of PAI‑1 in HepG2 cells via HIF‑1α." in: Molecular medicine reports, Vol. 14, Issue 2, pp. 1841-8, 2016 (PubMed).

Schokrpur, Hu, Moughon, Liu, Lin, Hermann, Mangul, Guan, Pellegrini, Xu, Wu: "CRISPR-Mediated VHL Knockout Generates an Improved Model for Metastatic Renal Cell Carcinoma." in: Scientific reports, Vol. 6, pp. 29032, 2016 (PubMed). Further details: Immunocytochemistry,Western Blotting,Immunofluorescence

Stei, Loeffler, Kurts, Hoeller, Pfarrer, Holz, Herwig-Carl: "Impact of macrophages on tumor growth characteristics in a murine ocular tumor model." in: Experimental eye research, Vol. 151, pp. 9-18, 2016 (PubMed). (Sample species: Human). Further details: Immunohistochemistry