Product: HIF1A Antibody
Catalog: AF1009
Source: Rabbit
Application: WB, IHC, IF/ICC, ELISA(peptide)
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Rabbit
Mol.Wt.: 120kD; 93kD(Calculated).
Uniprot: Q16665
RRID: AB_2835328

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:200, ELISA(peptide) 1:20000-1:40000
*The optimal dilutions should be determined by the end user.
Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Bovine(100%), Horse(100%), Rabbit(100%)
Clonality:
Polyclonal
Specificity:
HIF1A Antibody detects endogenous levels of total HIF1A.
RRID:
AB_2835328
Cite Format: Affinity Biosciences Cat# AF1009, RRID:AB_2835328.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.Stable for 15 months from date of receipt. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

ARNT interacting protein; ARNT-interacting protein; Basic helix loop helix PAS protein MOP1; Basic-helix-loop-helix-PAS protein MOP1; bHLHe78; Class E basic helix-loop-helix protein 78; HIF 1A; HIF 1alpha; HIF-1-alpha; HIF1 A; HIF1 Alpha; HIF1; HIF1-alpha; HIF1A; HIF1A_HUMAN; Hypoxia inducible factor 1 alpha; Hypoxia inducible factor 1 alpha isoform I.3; Hypoxia inducible factor 1 alpha subunit; Hypoxia inducible factor 1 alpha subunit basic helix loop helix transcription factor; Hypoxia inducible factor 1, alpha subunit (basic helix loop helix transcription factor); Hypoxia inducible factor1alpha; Hypoxia-inducible factor 1-alpha; Member of PAS protein 1; Member of PAS superfamily 1; Member of the PAS Superfamily 1; MOP 1; MOP1; PAS domain-containing protein 8; PASD 8; PASD8;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
Q16665 HIF1A_HUMAN:

Expressed in most tissues with highest levels in kidney and heart. Overexpressed in the majority of common human cancers and their metastases, due to the presence of intratumoral hypoxia and as a result of mutations in genes encoding oncoproteins and tumor suppressors. A higher level expression seen in pituitary tumors as compared to the pituitary gland.

Description:
Cell growth and viability is compromised by oxygen deprivation (hypoxia). Hypoxia-inducible factors, including HIF-1α, Arnt 1 (also designated HIF-1β), EPAS-1 (also designated HIF-2α) and HIF-3α, induce glycolysis, erythropoiesis and angiogenesis in order to restore oxygen homeostasis. Hypoxia-inducible factors are members of the Per-Arnt-Sim (PAS) domain transcription factor family.
Sequence:
MEGAGGANDKKKISSERRKEKSRDAARSRRSKESEVFYELAHQLPLPHNVSSHLDKASVMRLTISYLRVRKLLDAGDLDIEDDMKAQMNCFYLKALDGFVMVLTDDGDMIYISDNVNKYMGLTQFELTGHSVFDFTHPCDHEEMREMLTHRNGLVKKGKEQNTQRSFFLRMKCTLTSRGRTMNIKSATWKVLHCTGHIHVYDTNSNQPQCGYKKPPMTCLVLICEPIPHPSNIEIPLDSKTFLSRHSLDMKFSYCDERITELMGYEPEELLGRSIYEYYHALDSDHLTKTHHDMFTKGQVTTGQYRMLAKRGGYVWVETQATVIYNTKNSQPQCIVCVNYVVSGIIQHDLIFSLQQTECVLKPVESSDMKMTQLFTKVESEDTSSLFDKLKKEPDALTLLAPAAGDTIISLDFGSNDTETDDQQLEEVPLYNDVMLPSPNEKLQNINLAMSPLPTAETPKPLRSSADPALNQEVALKLEPNPESLELSFTMPQIQDQTPSPSDGSTRQSSPEPNSPSEYCFYVDSDMVNEFKLELVEKLFAEDTEAKNPFSTQDTDLDLEMLAPYIPMDDDFQLRSFDQLSPLESSSASPESASPQSTVTVFQQTQIQEPTANATTTTATTDELKTVTKDRMEDIKILIASPSPTHIHKETTSATSSPYRDTQSRTASPNRAGKGVIEQTEKSHPRSPNVLSVALSQRTTVPEEELNPKILALQNAQRKRKMEHDGSLFQAVGIGTLLQQPDDHAATTSLSWKRVKGCKSSEQNGMEQKTIILIPSDLACRLLGQSMDESGLPQLTSYDCEVNAPIQGSRNLLQGEELLRALDQVN

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Rabbit
100
Chicken
55
Sheep
0
Dog
0
Xenopus
0
Zebrafish
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q16665 As Substrate

Site PTM Type Enzyme
K10 Acetylation
K10 Sumoylation
K11 Acetylation
K12 Acetylation
K19 Acetylation
K21 Acetylation
S31 Phosphorylation
K32 Ubiquitination
T63 Phosphorylation
Y66 Phosphorylation
K71 Ubiquitination
K85 Ubiquitination
S113 Phosphorylation
K172 Ubiquitination
K185 Sumoylation
K185 Ubiquitination
S247 Phosphorylation P48730 (CSNK1D)
K251 Ubiquitination
Y276 Phosphorylation
K289 Ubiquitination
K297 Ubiquitination
K377 Ubiquitination
K389 Acetylation
K389 Ubiquitination
K391 Methylation
K391 Sumoylation
K391 Ubiquitination
S451 Phosphorylation
T455 Phosphorylation
T458 Phosphorylation
K460 Ubiquitination
S465 Phosphorylation
K477 Sumoylation
K477 Ubiquitination
C520 S-Nitrosylation
K532 Acetylation
K532 Ubiquitination
K538 Ubiquitination
K547 Ubiquitination
S551 Phosphorylation
T555 Phosphorylation
Y565 Phosphorylation
S576 Phosphorylation Q9H4B4 (PLK3)
S589 Phosphorylation
K636 Ubiquitination
S641 Phosphorylation P28482 (MAPK1) , P27361 (MAPK3)
S643 Phosphorylation P28482 (MAPK1) , P27361 (MAPK3)
K649 Ubiquitination
T651 Phosphorylation
T652 Phosphorylation
S653 Phosphorylation
S656 Phosphorylation
S657 Phosphorylation Q9H4B4 (PLK3)
S668 Phosphorylation P06493 (CDK1)
K674 Acetylation
K674 Ubiquitination
K682 Ubiquitination
S683 Phosphorylation
S687 Phosphorylation
S692 Phosphorylation
S696 Phosphorylation Q13315 (ATM)
T700 Phosphorylation
K709 Acetylation
K709 Ubiquitination
K721 Ubiquitination
S727 Phosphorylation
S760 Phosphorylation
S761 Phosphorylation
K769 Ubiquitination
T796 Phosphorylation
S797 Phosphorylation Q5S007 (LRRK2)
C800 S-Nitrosylation
S809 Phosphorylation

Research Backgrounds

Function:

Functions as a master transcriptional regulator of the adaptive response to hypoxia. Under hypoxic conditions, activates the transcription of over 40 genes, including erythropoietin, glucose transporters, glycolytic enzymes, vascular endothelial growth factor, HILPDA, and other genes whose protein products increase oxygen delivery or facilitate metabolic adaptation to hypoxia. Plays an essential role in embryonic vascularization, tumor angiogenesis and pathophysiology of ischemic disease. Heterodimerizes with ARNT; heterodimer binds to core DNA sequence 5'-TACGTG-3' within the hypoxia response element (HRE) of target gene promoters (By similarity). Activation requires recruitment of transcriptional coactivators such as CREBBP and EP300. Activity is enhanced by interaction with both, NCOA1 or NCOA2. Interaction with redox regulatory protein APEX seems to activate CTAD and potentiates activation by NCOA1 and CREBBP. Involved in the axonal distribution and transport of mitochondria in neurons during hypoxia.

PTMs:

S-nitrosylation of Cys-800 may be responsible for increased recruitment of p300 coactivator necessary for transcriptional activity of HIF-1 complex.

Requires phosphorylation for DNA-binding. Phosphorylation at Ser-247 by CSNK1D/CK1 represses kinase activity and impairs ARNT binding. Phosphorylation by GSK3-beta and PLK3 promote degradation by the proteasome.

Sumoylated; with SUMO1 under hypoxia. Sumoylation is enhanced through interaction with RWDD3. Both sumoylation and desumoylation seem to be involved in the regulation of its stability during hypoxia. Sumoylation can promote either its stabilization or its VHL-dependent degradation by promoting hydroxyproline-independent HIF1A-VHL complex binding, thus leading to HIF1A ubiquitination and proteasomal degradation. Desumoylation by SENP1 increases its stability amd transcriptional activity. There is a disaccord between various publications on the effect of sumoylation and desumoylation on its stability and transcriptional activity.

Acetylation of Lys-532 by ARD1 increases interaction with VHL and stimulates subsequent proteasomal degradation. Deacetylation of Lys-709 by SIRT2 increases its interaction with and hydroxylation by EGLN1 thereby inactivating HIF1A activity by inducing its proteasomal degradation.

Polyubiquitinated; in normoxia, following hydroxylation and interaction with VHL. Lys-532 appears to be the principal site of ubiquitination. Clioquinol, the Cu/Zn-chelator, inhibits ubiquitination through preventing hydroxylation at Asn-803. Ubiquitinated by a CUL2-based E3 ligase.

In normoxia, is hydroxylated on Pro-402 and Pro-564 in the oxygen-dependent degradation domain (ODD) by EGLN1/PHD2 and EGLN2/PHD1. EGLN3/PHD3 has also been shown to hydroxylate Pro-564. The hydroxylated prolines promote interaction with VHL, initiating rapid ubiquitination and subsequent proteasomal degradation. Deubiquitinated by USP20. Under hypoxia, proline hydroxylation is impaired and ubiquitination is attenuated, resulting in stabilization. In normoxia, is hydroxylated on Asn-803 by HIF1AN, thus abrogating interaction with CREBBP and EP300 and preventing transcriptional activation. This hydroxylation is inhibited by the Cu/Zn-chelator, Clioquinol. Repressed by iron ion, via Fe(2+) prolyl hydroxylase (PHD) enzymes-mediated hydroxylation and subsequent proteasomal degradation.

The iron and 2-oxoglutarate dependent 3-hydroxylation of asparagine is (S) stereospecific within HIF CTAD domains.

Subcellular Location:

Cytoplasm. Nucleus. Nucleus speckle.
Note: Colocalizes with HIF3A in the nucleus and speckles (By similarity). Cytoplasmic in normoxia, nuclear translocation in response to hypoxia (PubMed:9822602).

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Expressed in most tissues with highest levels in kidney and heart. Overexpressed in the majority of common human cancers and their metastases, due to the presence of intratumoral hypoxia and as a result of mutations in genes encoding oncoproteins and tumor suppressors. A higher level expression seen in pituitary tumors as compared to the pituitary gland.

Subunit Structure:

Interacts with the ARNT; forms a heterodimer that binds core DNA sequence 5'-TACGTG-3' within the hypoxia response element (HRE) of target gene promoters. Interacts with COPS5; the interaction increases the transcriptional activity of HIF1A through increased stability (By similarity). Interacts with EP300 (via TAZ-type 1 domains); the interaction is stimulated in response to hypoxia and inhibited by CITED2. Interacts with CREBBP (via TAZ-type 1 domains). Interacts with NCOA1, NCOA2, APEX and HSP90. Interacts (hydroxylated within the ODD domain) with VHLL (via beta domain); the interaction, leads to polyubiquitination and subsequent HIF1A proteasomal degradation. During hypoxia, sumoylated HIF1A also binds VHL; the interaction promotes the ubiquitination of HIF1A. Interacts with SENP1; the interaction desumoylates HIF1A resulting in stabilization and activation of transcription. Interacts (Via the ODD domain) with ARD1A; the interaction appears not to acetylate HIF1A nor have any affect on protein stability, during hypoxia. Interacts with RWDD3; the interaction enhances HIF1A sumoylation. Interacts with TSGA10 (By similarity). Interacts with HIF3A (By similarity). Interacts with RORA (via the DNA binding domain); the interaction enhances HIF1A transcription under hypoxia through increasing protein stability. Interaction with PSMA7 inhibits the transactivation activity of HIF1A under both normoxic and hypoxia-mimicking conditions. Interacts with USP20. Interacts with RACK1; promotes HIF1A ubiquitination and proteasome-mediated degradation. Interacts (via N-terminus) with USP19. Interacts with SIRT2. Interacts (deacetylated form) with EGLN1. Interacts with CBFA2T3. Interacts with HSP90AA1 and HSP90AB1.

Family&Domains:

Contains two independent C-terminal transactivation domains, NTAD and CTAD, which function synergistically. Their transcriptional activity is repressed by an intervening inhibitory domain (ID).

Research Fields

· Cellular Processes > Transport and catabolism > Autophagy - animal.   (View pathway)

· Environmental Information Processing > Signal transduction > HIF-1 signaling pathway.   (View pathway)

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Proteoglycans in cancer.

· Human Diseases > Cancers: Specific types > Renal cell carcinoma.   (View pathway)

· Human Diseases > Cancers: Overview > Central carbon metabolism in cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Choline metabolism in cancer.   (View pathway)

· Organismal Systems > Immune system > Th17 cell differentiation.   (View pathway)

· Organismal Systems > Endocrine system > Thyroid hormone signaling pathway.   (View pathway)

References

1). Zhong W et al. 6-Gingerol stabilized the p-VEGFR2/VE-cadherin/β-catenin/actin complex promotes microvessel normalization and suppresses tumor progression. J Exp Clin Cancer Res 2019 Jul 2;38(1):285 (PubMed: 31266540) [IF=7.068]

Application: IHC    Species: mouse    Sample: lung

Fig. 2 |6G suppresses the growth and metastasis, improves the tumor microenvironment and decreased MSE in vivo. a and (b) Representative tumors and in vivo bioluminescent images and tumor volume are shown. Each point represents the mean ± SD for different animal measurements (n = 10). *, P < 0.05;**, P < 0.01, one-way ANOVA. 6G inhibited TCs proliferation with dose-dependent manner. c Mice weight of each groups. dRepresentative lung metastasis specimens were sectioned and stained with h&e in 6G-treated group and control group. *, P < 0.05; **, P < 0.01, one-way ANOVA.

2). Zhang L et al. Estrogen stabilizes hypoxia-inducible factor 1α through G protein-coupled estrogen receptor 1 in eutopic endometrium of endometriosis. Fertil Steril 2017 Feb;107(2):439-447 (PubMed: 27939762) [IF=6.312]

Application: WB    Species: human    Sample:

17b-Estradiol and G1 regulate GPER and HIF-1a expression in ESCs. (A–D) Time course of GPER and HIF-1a mRNA levels in ESCs treated with 10 nM E2 or 100 nM G1 for 0, 5, 10, 15, 30, 60, and 120 minutes. (E, I) Time course of GPER and HIF-1a mRNA levels in ESCs treated with 10 nM E2 or 100 nM G1 for 0, 5, 10, 15, 30, 60, and 120 minutes. (F, G, J, K) Quantitative comparison of the fold difference in the expression of GPER and HIF- 1a proteins (*P<.05, ** P<.01, *** P<.001, R ¼ 0.7014) or G1 (P<.001, R ¼ 0.6386). (M–P) Time course of VEGF and MMP9 secretion after treatment with 10 nM E2 or 100 nM G1 for 0, 5, 10, 15, 30, 60, and 120 minutes.

Application: IHC    Species: human    Sample:

G protein-coupled estrogen receptor (GPER) and HIF-1a expression and localization in EuEM of endometriosis and CoEM. (A–D) Immunohistochemical analysis of GPER and HIF-1a protein expression and localization in CoEM. (E–H) Immunohistochemical analysis of GPER and HIF-1a protein in EuEM. Photographs were taken at original magnifications of 200 (left) and 400 (right), respectively. (I–L) Quantitative comparison of the fold difference in the expression of GPER and HIF-1a protein. The data are presented as means  SEM (*P<.05, ** P<.01, *** P

3). Xiong Y et al. Hypoxia-inducible factor 1α-induced epithelial-mesenchymal transition of endometrial epithelial cells may contribute to the development of endometriosis. Hum Reprod 2016 Jun;31(6):1327-38 (PubMed: 27094478) [IF=5.733]

Application: WB    Species: human    Sample:

Figure 2 The expression of epithelial –mesenchymal transition (EMT) markers and b-catenin was detected at each time point. (A) Immunoblotting analysis of human primary cultured endometrial epithelial cell extracts using the corresponding antibodies. The ratios of each protein relative to non-treated cells were normalized to GAPDH. (B) The relative expression of HIF-1a, N-cadherin, E-cadherin, b-catenin, vimentin and snail proteins in human endometrial epithelial glands under hypoxic conditions at each time point was investigated by western blot. Data are represented as mean+SD and are representative of the relative expression of protein normalized by GAPDH. All experiments were repeated four times. Data were evaluated by one-way ANOVA analysis (*P , 0.05, **P , 0.01 compared with untreated group). (C) The changed cellular morphologies of human endometrial epithelial glands in hypoxia compared with cells in normoxia, the hypoxic time was 48 h. Red arrows indicate the spindle-shaped and fibroblast-like cells.

Application: IHC    Species: human    Sample:

Figure 1 EMT occurs in endometrial epithelial cells of ovarian endometriosis samples. Representative photomicrographs of HIF-1a (A–C), b-catenin (D–F), E-cadherin (G–I), N-cadherin (J–L) and vimentin (M–O) in normal endometrium (A, D, G, J, M), eutopic endometrium (B, E, H, K, N) and ovarian endometriosis (C, F, I, L,O). (P)Colon cancer tissue that was positive for HIF-1a. (Q) Healthy liver tissue that was negative for HIF-1a. (R) Peptide-blocking reagent without antibody was applied as the negative controls. Photographs were taken at magnifications of ×200 (left panels) and ×400 (right panels). N, normal endometrium; U, eutopic endometrium; E, ovarian endometriosis.

4). Chen C et al. A multifunctional-targeted nanoagent for dual-mode image-guided therapeutic effects on ovarian cancer cells. Int J Nanomedicine 2019 Jan 21;14:753-769 (PubMed: 30718954) [IF=5.115]

Application: WB    Species: human    Sample: SKOV3 cells

Figure 6 |The underlying mechanism of therapy.(D, E) HIF-1α and IL-6 protein expression in SKOV3 cells was analyzed by Western blot after different treatments. GAPDH was used as an internal reference. Compared with control, *P,0.05; compared with other groups, #P,0.05.

5). Guo Y et al. The anti-dysenteric drug fraxetin enhances anti-tumor efficacy of gemcitabine and suppresses pancreatic cancer development by antagonizing STAT3 activation. Aging (Albany NY) 2021 Jul 28;13(14):18545-18563. (PubMed: 34320467) [IF=4.831]

6). Liu M et al. Luteolin alleviates ochratoxin A induced oxidative stress by regulating Nrf2 and HIF-1α pathways in NRK-52E rat kidney cells. Food Chem Toxicol 2020 May 18;111436. (PubMed: 32439592) [IF=4.679]

7). Tu J et al. Improving Tumor Hypoxia and Radiotherapy Resistance via in situ Nitric Oxide Release Strategy. Eur J Pharm Biopharm 2020 Mar 6 (PubMed: 32151726) [IF=4.604]

Application: WB    Species: Mouse    Sample: LLC tumor tissue

Fig. 4. TPGS-NO can effectively reduce hypoxia and promote angiogenesis. (a) IF staining analysis was employed to detect the CD31 (red) and Glut-1 (green) in LLC tumor tissue after different treatment (PBS, TPGS-NO, RT (8 GY), TPGS-NO + RT (8 GY), scale bar for 200 µm). IF quantitative analysis of (b) CD31 and (c) Glut-1. (d) Expression of CD31 and HIF-1α analyzed by Western blot. (e) and (f) are quantitative statistics of Western blot. Data are given as mean ± SD (n = 3). ** p < 0.01, ***p < 0.001. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

8). Wang X et al. Curcumin pretreatment protects against hypoxia/reoxgenation injury via improvement of mitochondrial function, destabilization of HIF-1α and activation of Epac1-Akt pathway in rat bone marrow mesenchymal stem cells. Biomed Pharmacother 2019 Jan;109:1268-1275 (PubMed: 30551377) [IF=4.545]

9). Sun Z et al. Long non-coding RNA and mRNA profile analysis of metformin to reverse the pulmonary hypertension vascular remodeling induced by monocrotaline. Biomed Pharmacother 2019 May 3;115:108933 (PubMed: 31060005) [IF=4.545]

10). Gong Q et al. Enhanced ROBO4 is mediated by up-regulation of HIF-1α/SP1 or reduction in miR-125b-5p/miR-146a-5p in diabetic retinopathy. J Cell Mol Med 2019 May 15 (PubMed: 31094072) [IF=4.486]

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