Anti-HIF-1 alpha antibody [mgc3]

• 产品名称

  Anti-HIF-1 alpha抗体[mgc3]
  参阅全部 HIF-1 alpha 一抗

• 描述

  小鼠单克隆抗体[mgc3] to HIF-1 alpha

• 宿主

  Mouse

• 特异性

  This antibody does not cross-react with ARNT or the related HIF-2-alpha.

• 经测试应用

  适用于: Flow Cyt, WB, IHC-Pmore details

• 种属反应性

  与反应: Human
  预测可用于: Mouse, Cow, Pig, Non human primates

• 免疫原

  Recombinant fragment corresponding to Human HIF-1 alpha aa 530-826 (C terminal).

• 阳性对照

• IHC-P: Human colon tissue. Human small intestine and tonsil tissue. Flow Cyt: HeLa cells. WB: HeLa wild type treated with 150uM CoCl2 for 48 hrs and HeLa Cas9 treated with 150uM CoCl2 for 48 hrs.

• 常规说明

  Under normoxic conditions HIF-1 alpha has a short half-life. It is largely undetectable in cells or tissues grown under normoxic conditions. It is stabilized only at O2 concentrations below 5% and upon stabilization under hypoxic conditions HIF-1 translocates to the nucleus. Hypoxia can be induced with treatment using certain agents e.g. CoCl₂ or DFO, etc. so proper sample preparation is critical.

  
  

  The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

  If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

• 形式

  Liquid

• 存放说明

  Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.

• 存储溶液

  pH: 7.4
  Preservative: 0.05% Sodium azide
  Constituent: 0.1% BSA

• 浓度

• 100 µl 浓度为 1 mg/ml

• 纯度

  Protein G purified

• 克隆

  单克隆

• 克隆编号

  mgc3

• 同种型

  IgG1

The Abpromise guarantee

Abpromise™承诺保证使用ab16066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。

• 数据库链接

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• Unigene: 3879 Mouse

• Unigene: 446610 Mouse

• Entrez Gene: 281814 Cow

• Entrez Gene: 3091 Human

• Entrez Gene: 15251 Mouse

• Entrez Gene: 396696 Pig

• Omim: 603348 Human

• SwissProt: Q16665 Human

• SwissProt: Q61221 Mouse

• Unigene: 597216 Human

• 别名

• Basic-helix-loop-helix-PAS protein MOP1 antibody

• bHLHe78 antibody

• Class E basic helix-loop-helix protein 78 antibody

• HIF 1A antibody

• HIF 1alpha antibody

• HIF-1-alpha antibody

• HIF-1alpha antibody

• HIF-alpha antibody

• HIF1 A antibody

• HIF1 Alpha antibody

• HIF1 antibody

• HIF1-alpha antibody

• HIF1A antibody

• HIF1A_HUMAN antibody

• hifla antibody

• Hypoxia inducible factor 1 alpha antibody

• Hypoxia inducible factor 1 alpha isoform I.3 antibody

• Hypoxia inducible factor 1 alpha subunit antibody

• Hypoxia inducible factor 1 alpha subunit basic helix loop helix transcription factor antibody

• Hypoxia inducible factor 1, alpha subunit (basic helix loop helix transcription factor) antibody

• Hypoxia inducible factor1alpha antibody

• Hypoxia-inducible factor 1-alpha antibody

• Member of PAS protein 1 antibody

• Member of PAS superfamily 1 antibody

• Member of the PAS Superfamily 1 antibody

• MOP 1 antibody

• MOP1 antibody

• PAS domain-containing protein 8 antibody

• PASD 8 antibody

• PASD8 antibody

• ARNT interacting protein antibody

• ARNT-interacting protein antibody

• Basic helix loop helix PAS protein MOP1 antibody

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF-1 alpha antibody [mgc3] (ab16066)

  IHC image of ab16066 staining HIF-1 alpha in human colon formalin fixed paraffin embedded tissue sections*, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH9, epitope retrieval solution 2) for 20 mins. The section was then incubated with ab16066, 10μg/ml working concentration, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).
  For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
  *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• 

  Flow Cytometry - Anti-HIF-1 alpha antibody [mgc3] (ab16066)

  Overlay histogram showing HeLa cells stained with ab16066 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab16066, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF-1 alpha antibody [mgc3] (ab16066)This image is courtesy of an anonymous Abreview

  ab16066 staining HIF-1-alpha in Human small intestine (IBD) and tonsil tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde; antigen retrieval was by heat mediation with an EDTA buffer (pH 9.0). Samples were incubated with primary antibody (1/800 in diluent + background reducers) for 20 minutess at 25°C. An undiluted Goat polymer was used as the secondary antibody.

  See Abreview

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF-1 alpha antibody [mgc3] (ab16066)

  Immunohistochemistry was performed on normal biopsies of deparaffinized Human tonsil tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1/20 with ab16066 (left) or without primary antibody (negative control - right) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

• 

  Western blot - Anti-HIF-1 alpha antibody [mgc3] (ab16066)

  All lanes : Anti-HIF-1 alpha antibody [mgc3] (ab16066) at 1/2000 dilution
  
  Lane 1 : HeLa wild type (untreated)
  Lane 2 : HeLa wild type treated with 150uM CoCl2 for 48 hrs
  Lane 3 : HeLa Cas9
  Lane 4 : HeLa Cas9 treated with 150uM CoCl2 for 48 hrs
  Lane 5 : HeLa HIF1A KO
  Lane 6 : HeLa HIF1A KO treated with 150uM CoCl2 for 48 hrs
  
  Lysates/proteins at 30 µg per lane.
  
  Secondary
  All lanes : Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP at 1/4000 dilution
  
  Predicted band size: 92 kDa
  Observed band size: 90 kDawhy is the actual band size different from the predicted?
  
  
  
  

  Antibody specificity was demonstrated by CRISPR-Cas9 mediated knockout of target protein. A loss of signal was observed for target protein in HIF-1 alpha KO cell line compared to control cell lines using ab16066. 

  Uncharacterized bands were observed in HeLa Cas9 samples at ~40 kDa and 45 kDa.