Anti-BBC3/PUMA antibody

Anti-BBC3/PUMA antibody

• 概述

• 产品描述Essential mediator of p53/TP53-dependent and p53/TP53-independent apoptosis. Functions by promoting partial unfolding of BCL2L1 and dissociation of BCL2L1 from p53/TP53. Essential for ER stress-induced cell death.
• 产品名称Anti-BBC3/PUMA antibody
• 分子量21 KDa
• 种属反应性Human,Mouse,Rat,Dog,Pig,Cow
• 验证应用WB,IHC-P,FC
• 抗体类型兔多抗
• 免疫原KLH conjugated synthetic peptide derived from human BBC3 131-180/193
• 偶联Non-conjugated

• 性能

• 形态Liquid
• 浓度1 mg/mL.
• 存放说明Store at -20℃ for one year. Avoid repeated freeze/thaw cycles. The lyophilized antibody is stable at room temperature for at least one month and for greater than a year when kept at -20℃. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4℃.
• 存储缓冲液0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol.
• 亚型IgG
• 纯化方式affinity purified by Protein A
• 亚细胞定位Mitochondrion. Note=Localized to the mitochondria in order to induce cytochrome c release.
• 其它名称PUMA
  bcl-2 binding component 3
  BBC 3
  BBC3
  BCL 2 binding component 3
  BCL2 binding component 3
  JFY 1
  JFY1
  p53 up regulated modulator of apoptosis
  p53 Upregulated Modulator of Apoptosis
  PUMA alpha
  PUMA/JFY1
  BBC3_RAT.
  more

• 应用

  WB:1:500-2000
  IHC-P:1:400-800
  FC:1μg /test

•  

  

  Fig1: Tissue/cell: rat brain tissue; 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-BBC3/PUMA Polyclonal Antibody, Unconjugated1:200, overnight at 4℃, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

   

  

  Fig2: Blank control: U-87MG(blue).
  Primary Antibody:Rabbit Anti-BBC3 PUMA antibody Dilution: 1μg in 100 μL 1X PBS containing 0.5% BSA;
  Isotype Control Antibody: Rabbit IgG(orange) ,used under the same conditions );
  Secondary Antibody: Goat anti-rabbit IgG-PE(white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.
  Protocol
  The cells were fixed with 2% paraformaldehyde (10 min) , then permeabilized with 90% ice-cold methanol for 30 min on ice. Primary antibody (ER1904-14,1μg /1x10^6 cells) were incubated for 30 min on the ice, followed by 1 X PBS containing 0.5% BSA + 1 0% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/PE antibody was added into the blocking buffer mentioned above to react with the primary antibody at 1/200 dilution for 30 min on ice. Acquisition of 20,000 events was performed.

   

  

  Fig3: Sample: K562 Cell Lysate at 30 ug
  Primary: Anti- BBC3 at 1/300 dilution
  Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
  Predicted band size: 21 kD
  Observed band size: 25 kD

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