小鼠HIF-1 alpha ELISA试剂盒(ab275103)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 33.39 pg/ml
- Range: 117.19 pg/ml - 7500 pg/ml
- Sample type: Cell Lysate, Tissue Lysate
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Mouse
概述
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产品名称
小鼠HIF-1 alpha ELISA试剂盒
参阅全部 HIF-1 alpha 试剂盒 -
检测方法
Colorimetric -
精确度
批次内 样品 n Mean SD CV% Extract 8 10% 批次间 样品 n Mean SD CV% Extract 3 15% -
样品类型
Cell Lysate, Tissue Lysate -
检测类型
Sandwich (quantitative) -
灵敏度
33.39 pg/ml -
范围
117.19 pg/ml - 7500 pg/ml -
回收率
特定样本回收率 样品类型 平均% 范围 Cell Lysate 96 87% - 106% -
检测时间
1h 30m -
实验步骤
One step assay -
种属反应性
与反应: Mouse -
产品概述
Mouse HIF-1 alpha ELISA kit (ab275103) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Mouse HIF-1 alpha protein in cell and tissue extracts. It uses our proprietary SimpleStep ELISA® technology. Quantitate Mouse HIF-1 alpha with 33.39 pg/mL sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
-Single-wash protocol reduces assay time to 90 minutes or less
-High sensitivity, specificity and reproducibility from superior antibodies
-Fully validated in biological samples
-96-wells plate breakable into 12 x 8 wells strips
A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
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说明
Hypoxia-inducible factor 1-alpha (HIF-1 alpha) is a constitutively expressed transcription factor that is degraded under normal oxygen tensions and stabilized when oxygen is limiting (hypoxia). Under hypoxic conditions, stabilized HIF-1 alpha translocates to the nucleus and promotes the transcription of genes that enable the cell to adapt to a low oxygen environment. The HIF-1 alpha mediated hypoxic response include promotion of angiogenesis and the switch from aerobic respiration to anaerobic glycolysis. Many of the HIF-1 alpha responsive genes encode proteins that promote glycolysis and/or inhibit oxidative phosphorylation included in the Warburg effect.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses. -
经测试应用
适用于: Sandwich ELISAmore details -
平台
Pre-coated microplate (12 x 8 well strips)
性能
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存放说明
Store at +4°C. Please refer to protocols. -
组件 1 x 96 tests 10X Mouse HIF-1 alpha Capture Antibody 1 x 600µl 10X Mouse HIF-1 alpha Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent 5BR 1 x 6ml Mouse HIF-1 alpha Lyophilized Recombinant Protein 2 vials Plate Seal 1 unit Sample Diluent NS (ab193972) 1 x 12ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml -
研究领域
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功能
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, 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. Binds to core DNA sequence 5'-[AG]CGTG-3' within the hypoxia response element (HRE) of target gene promoters. Activation requires recruitment of transcriptional coactivators such as CREBPB 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. -
组织特异性
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. -
序列相似性
Contains 1 basic helix-loop-helix (bHLH) domain.
Contains 1 PAC (PAS-associated C-terminal) domain.
Contains 2 PAS (PER-ARNT-SIM) domains. -
结构域
Contains two independent C-terminal transactivation domains, NTAD and CTAD, which function synergistically. Their transcriptional activity is repressed by an intervening inhibitory domain (ID). -
翻译后修饰
In normoxia, is hydroxylated on Pro-402 and Pro-564 in the oxygen-dependent degradation domain (ODD) by EGLN1/PHD1 and EGLN2/PHD2. 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.
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.
Sumoylated; by SUMO1 under hypoxia. Sumoylation is enhanced through interaction with RWDD3. Desumoylation by SENP1 leads to increased HIF1A stability and transriptional activity.
Ubiquitinated; 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.
The iron and 2-oxoglutarate dependent 3-hydroxylation of asparagine is (S) stereospecific within HIF CTAD domains. -
细胞定位
Cytoplasm. Nucleus. Cytoplasmic in normoxia, nuclear translocation in response to hypoxia. Colocalizes with SUMO1 in the nucleus, under hypoxia. - Information by UniProt
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别名
- ARNT interacting protein
- ARNT-interacting protein
- Basic helix loop helix PAS protein MOP1
see all -
数据库链接
- Entrez Gene: 15251 Mouse
- SwissProt: Q61221 Mouse
- Unigene: 3879 Mouse
- Unigene: 446610 Mouse
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab275103于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| Sandwich ELISA |
Use at an assay dependent concentration.
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| 说明 |
|---|
|
Sandwich ELISA
Use at an assay dependent concentration. |
图片
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SimpleStep ELISA Protocol Diagram
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
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Example of mouse HIF-1alpha standard curve in 1X Cell Extraction Buffer PTR.The HIF-1alpha standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of native HIF-1alpha in mouse NIH/3T3 cell extract either treated with deferoxamine or mock treatment based on a 100 µg/mL extract load.The concentrations of HIF-1alpha were measured in duplicate, interpolated from the HIF-1alpha standard curves and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean HIF-1alpha concentration was determined to be 1481 pg/mL.
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Interpolated concentrations of native HIF-1alpha in mouse deferoxamine treated NIH/3T3 cell extract based on a 10 µg/mL extract load.The concentrations of HIF-1alpha were measured in duplicate, interpolated from the HIF-1alpha standard curves and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean HIF-1alpha concentration was determined to be 485.55 pg/mL and 40.77 pg/ml in deferoxamine and mock treated cell extract, respectively.
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Sandwich ELISA - Mouse HIF-1 alpha ELISA Kit (ab275103)To learn more about the advantages of recombinant antibodies see here.
数据表及文件
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SDS download
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Datasheet download
文献 (1)
ab275103 被引用在 1 文献中.
- Abd El-Fattah EE et al. AKT-AMPKα-mTOR-dependent HIF-1α Activation is a New Therapeutic Target for Cancer Treatment: A Novel Approach to Repositioning the Antidiabetic Drug Sitagliptin for the Management of Hepatocellular Carcinoma. Front Pharmacol 12:720173 (2021). PubMed: 35095479