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-Yeast Hypoxia Inducible Factor 1, alpha Subunit (Basic Helix-Loop-Helix Transcription Factor) antibody for Chromatin Immunoprecipitation
Reactivity
Cow (Bovine), Ferret, Human, Monkey, Mouse (Murine), Pig (Porcine), Primate, Rabbit, Rat (Rattus), Sheep (Ovine), Xenopus laevis, Yeast
611
303
250
145
91
76
66
64
58
54
44
36
28
21
20
19
19
18
17
9
3
2
2
2
1
1
1
1
1
Host
Mouse
385
278
2
Clonality (Clone)
Monoclonal ()
Conjugate
Un-conjugated
52
33
33
26
25
25
18
18
15
14
12
12
6
6
6
4
4
4
4
3
3
3
2
2
2
2
2
2
2
2
2
2
2
2
1
1
1
1
1
1
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)
512
226
205
181
174
169
114
91
62
16
16
13
13
13
10
7
6
5
4
3
2
2
2
1
1
Options
Supplier
Log in to see
Supplier Product No.
Log in to see
Request

Get this product for free

It's quick and easy to submit your validation proposal. I want to validate this product

Learn more

Available images

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, Tube Formation, Regulation of Carbohydrate Metabolic Process, Signaling Events mediated by VEGFR1 and VEGFR2, VEGFR1 Specific Signals
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: Ponente, Campanini, Cuttano, Piunti, Delledonne, Coltella, Valsecchi, Villa, Cavallaro, Pattini, Doglioni, Bernardi: "PML promotes metastasis of triple-negative breast cancer through transcriptional regulation of HIF1A target genes." in: JCI insight, Vol. 2, Issue 4, pp. e87380, 2017 (PubMed). (Sample species: Human).

Qing, Lei, Liu, Xie, Wang, Wen, Hu: "Expression of hypoxia-inducible factor-1α in synovial fluid and articular cartilage is associated with disease severity in knee osteoarthritis." in: Experimental and therapeutic medicine, Vol. 13, Issue 1, pp. 63-68, 2017 (PubMed). Method employed by authors: Immunohistochemistry (IHC) (Sample species: Human).

Jeong, Jung, Cho, Park, Oh, Kang: "Correlation of hypoxia inducible transcription factor in breast cancer and SUVmax of F-18 FDG PET/CT." in: Nuclear medicine review. Central & Eastern Europe, Vol. 20, Issue 1, pp. 32-38, 2017 (PubMed). Method employed by authors: Immunohistochemistry (IHC) (Sample species: Human).

Yang, Yang, Wang, Kuang: "The hypoxia-inducible factors HIF1α and HIF2α are dispensable for embryonic muscle development but essential for postnatal muscle regeneration." in: The Journal of biological chemistry, Vol. 292, Issue 14, pp. 5981-5991, 2017 (PubMed). Method employed by authors: Western Blotting (WB) (Sample species: Human).

Mi, Ma, Wang, Zuo, Wang, Li, Piao, Xu, Li, Quan, Jin: "Imperatorin suppresses proliferation and angiogenesis of human colon cancer cell by targeting HIF-1α via the mTOR/p70S6K/4E-BP1 and MAPK pathways." in: Journal of ethnopharmacology, Vol. 203, pp. 27-38, 2017 (PubMed). Method employed by authors: Western Blotting (WB) (Sample species: Human).

Revenco, Lapouge, Moers, Brohée, Sotiropoulou: "Low Dose Radiation Causes Skin Cancer in Mice and Has a Differential Effect on Distinct Epidermal Stem Cells." in: Stem cells (Dayton, Ohio), Vol. 35, Issue 5, pp. 1355-1364, 2017 (PubMed). Method employed by authors: Immunofluorescence (fixed cells) (IF/ICC) (Sample species: Mouse (Murine)).

Ridiandries, Bursill, Tan: "Broad-Spectrum Inhibition of the CC-Chemokine Class Improves Wound Healing and Wound Angiogenesis." in: International journal of molecular sciences, Vol. 18, Issue 1, 2017 (PubMed). Method employed by authors: Western Blotting (WB) (Sample species: Mouse (Murine)).

Feng, Zhang, Lum, Druso, Blank, Wilson, Welm, Antonyak, Cerione: "A class of extracellular vesicles from breast cancer cells activates VEGF receptors and tumour angiogenesis." in: Nature communications, Vol. 8, pp. 14450, 2017 (PubMed). Method employed by authors: Western Blotting (WB) (Sample species: Mouse (Murine)).

Chung, Lin, Lee, Lee, Lee, Yang: "Brain-Derived Neurotrophic Factor Loaded PS80 PBCA Nanocarrier for In Vitro Neural Differentiation of Mouse Induced Pluripotent Stem Cells." in: International journal of molecular sciences, Vol. 18, Issue 3, 2017 (PubMed). Method employed by authors: Western Blotting (WB) (Sample species: Mouse (Murine)).

Takahara, Tokunou, Kojima, Hirooka, Ichiki: "Deletion of hypoxia-inducible factor-1α in myeloid lineage exaggerates angiotensin II-induced formation of abdominal aortic aneurysm." in: Clinical science (London, England : 1979), Vol. 131, Issue 7, pp. 609-620, 2017 (PubMed). (Sample species: Mouse (Murine)).

Fraga, Ribeiro, Coelho, Vizcaíno, Coutinho, Lopes, Príncipe, Lobato, Lopes, Medeiros: "Genetic polymorphisms in key hypoxia-regulated downstream molecules and phenotypic correlation in prostate cancer." in: BMC urology, Vol. 17, Issue 1, pp. 12, 2017 (PubMed).

Li, Zhou, Dai: "Retinoic acid receptor-related orphan receptor RORα regulates differentiation and survival of keratinocytes during hypoxia." in: Journal of cellular physiology, 2017 (PubMed).

An, Sun, Joffin, Zhang, Deng, Donzé, Kusminski, Scherer: "Angiopoietin-2 in white adipose tissue improves metabolic homeostasis through enhanced angiogenesis." in: eLife, Vol. 6, 2017 (PubMed).

Robb, Cotechini, Allaire, Sperou, Graham: "Inflammation-induced fetal growth restriction in rats is associated with increased placental HIF-1α accumulation." in: PLoS ONE, Vol. 12, Issue 4, pp. e0175805, 2017 (PubMed). Method employed by authors: Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)) (Sample species: Rat (Rattus)).

Yamazaki, Kasuya, Fujita, Umezawa, Yanagihara, Nakamura, Yoshino, Tatsumi, Murayama: "Antifibrotic effects of cyclosporine A on TGF-β1-treated lung fibroblasts and lungs from bleomycin-treated mice: role of hypoxia-inducible factor-1α." in: FASEB journal : official publication of the Federation of American Societies for Experimental Biology, Vol. 31, Issue 8, pp. 3359-3371, 2017 (PubMed).

Wu, Qian, Zhou, Yan, Luo, Yung, Zhang, Jiang, Qian, Ke: "Bi-directionally protective communication between neurons and astrocytes under ischemia." in: Redox biology, Vol. 13, pp. 20-31, 2017 (PubMed). Method employed by authors: Western Blotting (WB) (Sample species: Rat (Rattus)).

Fukushima, Endo, Matsumoto, Fukushi, Matsunobu, Kawaguchi, Setsu, IIda, Yokoyama, Nakagawa, Yahiro, Oda, Iwamoto, Nakashima: "Hypoxia-inducible factor 1 alpha is a poor prognostic factor and potential therapeutic target in malignant peripheral nerve sheath tumor." in: PLoS ONE, Vol. 12, Issue 5, pp. e0178064, 2017 (PubMed). Method employed by authors: Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)) (Sample species: Human).

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