| Rabbit Anti-HIF-1 Alpha antibody |
| 反应物种（预测） |
Mouse,Dog,Pig,Cow,Rabbit |
| 产品应用（已验证） |
WB,IHC,ICC,FCM |
| 产品应用（可尝试） |
IF,ELISA |
| 推荐稀释比例 |
WB=1:500-2000,Elisa=1:5000-10000,IHC-P=1:100-500,IHC-F=1:100-500,Flow Cyt=1μg/Test,IF=1:100-500,ICC=1:100, |
| 研究领域 |
肿瘤,细胞生物,神经生物学,信号转导,细胞凋亡 |
| 标签 |
Array |
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Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (HIF-1 Alpha) polyclonal Antibody, Unconjugated (bs-0737R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
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Paraformaldehyde-fixed, paraffin embedded (rat kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (HIF-1 Alpha) Polyclonal Antibody, Unconjugated (bs-0737R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
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Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (HIF-1 Alpha) polyclonal Antibody, Unconjugated (bs-0737R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
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Sample:
Lane 1: U87MG (Human) Cell Lysate at 30 ug
Lane 2: Hela (Human) Cell Lysate at 30 ug
Lane 3: A431 (Human) Cell Lysate at 30 ug
Lane 4: U251 (Human) Cell Lysate at 30 ug
Primary: Anti-HIF-1 Alpha (bs-0737R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 120 kD
Observed band size: 120 kD
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Paraformaldehyde-fixed, paraffin embedded (rat stomach); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (HIF-1 Alpha) Polyclonal Antibody, Unconjugated (bs-0737R ) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
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Blank control:Mouse spleen.
Primary Antibody (green line): Rabbit Anti-HIF-1 Alpha antibody (bs-0737R)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-FITC
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
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Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (HIF-1 Alpha) polyclonal Antibody, Unconjugated (bs-0737R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
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Blank control (blue line): Hela (blue).
Primary Antibody (green line): Rabbit Anti- HIF-1 Alpha antibody (bs-0737R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC
Dilution: 1μg /test.
Protocol
The cells were fixed with 80% methanol (5 min at -20℃) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2%BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
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Tissue/cell: rat lung tissue(Smoking); 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-HIF-1-Alpha Polyclonal Antibody, Unconjugated(bs-0737R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
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Tissue/cell: human cervical carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-HIF-1-Alpha Polyclonal Antibody, Unconjugated(bs-0737R) 1:300, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
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Sample:
Lane 1: Hela (Human) Cell Lysate at 30 ug
Lane 2: A431 (Human) Cell Lysate at 30 ug
Lane 3: U251 (Human) Cell Lysate at 30 ug
Lane 4: HepG2 (Human) Cell Lysate at 30 ug
Primary: Anti-HIF-1 Alpha (bs-0737R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 120 kD
Observed band size: 120 kD
RRID：AB_10857933
产品名称：Rabbit Anti-HIF-1 Alpha antibody
别名： 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;
中文名称：缺氧诱导因子1α /低氧诱导因子-1抗体
英文名称：Rabbit Anti-HIF-1 Alpha antibody
中文别名：HIF-1a; HIF-1α;
抗体来源： Rabbit
克隆类型：多克隆
细胞定位：细胞核,细胞浆
性 状：Liquid
亚 型：IgG
纯化方法：affinity purified by Protein A
保存条件：Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.
免 疫 原：KLH conjugated synthetic peptide derived from middle of human HIF-1 Alpha
抗原表位：341-450/826
SWISS：Q16665
Gene ID ：3091
Human Gene ID：3091
Hypoxia-inducible factor-1 (HIF1) is a transcription factor found in mammalian cells cultured under reduced oxygen tension that plays an essential role in cellular and systemic homeostatic responses to hypoxia. HIF1 is a heterodimer composed of an alpha subunit and a beta subunit. The beta subunit has been identified as the aryl hydrocarbon receptor nuclear translocator(ARNT). This gene encodes the alpha subunit of HIF-1. Overexpression of a natural antisense transcript (aHIF) of this gene has been shown to be associated with nonpapillary renal carcinomas. Two alternative transcripts encoding different isoforms have been identified.
Function：Functions as a master transcriptional regulator of theadaptive response to hypoxia. Under hypoxic conditions activatesthe transcription of over 40 genes, including, erythropoietin,glucose transporters, glycolytic enzymes, vascular endothelialgrowth factor
Subunit：Interacts with COPS5 subunit of COP9 signalosome complex,leading to the regulation of its stability. Interacts with TSGA10(By similarity). Efficient DNA binding requires heterodimerizationof an alpha and a beta/ARNT subunit. Binds to the TAZ-type 1domains
Subcellular Location：Cytoplasm. Nucleus.
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 tumo
Post-translational modifications：In normoxia, is hydroxylated on Pro-402 and Pro-564 in theoxygen-dependent degradation domain (ODD) by EGLN1/PHD1 andEGLN2/PHD2. EGLN3/PHD3 has also been shown to hydroxylate Pro-564.The hydroxylated prolines promote interaction with VHL, initiatingrapid
Similarity：Contains 1 basic helix-loop-helix (bHLH) domain.
Contains 1 PAC (PAS-associated C-terminal) domain.
Contains 2 PAS (PER-ARNT-SIM) domains.
Important Note：This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
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