Anti-Phosphotyrosine antibody [PY20]

• 产品名称

  Anti-Phosphotyrosine抗体[PY20]
  参阅全部 Phosphotyrosine 一抗

• 描述

  小鼠单克隆抗体[PY20] to Phosphotyrosine

• 宿主

  Mouse

• 经测试应用

  适用于: WB, ICC/IFmore details
  不适用于: IHC-P

• 种属反应性

  与反应: Species independent
  

• 免疫原

  Chemical/ Small Molecule corresponding to Phosphotyrosine conjugated to keyhole limpet haemocyanin.

• 阳性对照

• ICC/IF: C2C12 cells treated with 2mM H2O2 for 10min. WB: NIH 3T3 treated with Vanadate and PDGF Whole Cell Lysate.

• 常规说明

  
  

  This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

  
  

  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 or -80°C. Avoid freeze / thaw cycle.

• 存储溶液

  pH: 7.40
  Preservative: 0.02% Sodium azide
  Constituents: PBS, 6.97% L-Arginine
  
  Contains 0.4M Arginine

• 浓度

• 批次浓度范围 100 µg 浓度为 0.9 - 1 mg/ml

• 纯度

  Affinity purified

• Primary antibody说明

  This is a standard clone used to detect phosphotyrosine.

• 克隆

  单克隆

• 克隆编号

  PY20

• 同种型

  IgG2b

The Abpromise guarantee

Abpromise™承诺保证使用ab10321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。

应用说明

Is unsuitable for IHC-P.

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Phosphotyrosine antibody [PY20] (ab10321)

  ab10321 staining Phosphotyrosine in C2C12 cells treated with 2mM H2O2 for 10mins. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab10321 at 1µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

  Also suitable in cells fixed with 100% methanol (5 min).

  Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• 

  Western blot - Anti-Phosphotyrosine antibody [PY20] (ab10321)

  All lanes : Anti-Phosphotyrosine antibody [PY20] (ab10321) at 1 µg/ml
  
  Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
  Lane 2 : NIH 3T3 treated with Vanadate and PDGF Whole Cell Lysate
  
  Lysates/proteins at 5 µg per lane.
  
  Secondary
  All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
  
  Developed using the ECL technique.
  
  Performed under reducing conditions.
  
  Exposure time: 1 minute
  
  
  

  Cells were serum starved overnight and then incubated at room temperature for 10mins in a final concentration of 1mM sodium vanadate. PDGF was then added at a final concentration of 5ng/ml and cells were incubated at 37ºC for 30mins. Vanadate inhibits endogenous phosphatases and PDGF stimulates phosphorylation. Western blots of NIH 3T3 cell lysates treated with vanadate and PDGF show an array of phosphorylated tyrosine compared to controls.