Anti-Cyclin E1 antibody

Anti-Cyclin E1 antibody

• 概述

• 产品描述The protein encoded by this gene belongs to the highly conserved cyclin family, whose members are characterized by a dramatic periodicity in protein abundance through the cell cycle. Cyclins function as regulators of CDK. Different cyclins exhibit distinct expression and degradation patterns which contribute to the temporal coordination of each mitotic event. This cyclin forms a complex with and functions as a regulatory subunit of CDK2, whose activity is required for cell cycle G1/S transition. This protein accumulates at the G1-S phase boundary and is degraded as cells progress through S phase. Overexpression of this gene has been observed in many tumors, which results in chromosome instability, and thus may contribute to tumorigenesis. This protein was found to associate with, and be involved in, the phosphorylation of NPAT protein (nuclear protein mapped to the ATM locus), which participates in cell-cycle regulated histone gene expression and plays a critical role in promoting cell-cycle progression in the absence of pRB.
• 产品名称Anti-Cyclin E1 antibody
• 分子量45 KDa
• 种属反应性Human,Mouse,Rat
• 验证应用WB,IHC-P,ICC/IF,FC
• 抗体类型兔多抗
• 免疫原KLH conjugated synthetic peptide derived from rat Cyclin E 375-411/411
• 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Nucleus.
  
• 其它名称CCNE 1
  CCNE
  CCNE1
  Cyclin Es
  Cyclin Et
  CyclinE
  G1/S specific cyclin E
  G1/S-specific cyclin-E1
  CCNE1_HUMAN
  pCCNE1.
  more

• 应用

  WB:1:500-2000
  IHC-P:1:400-800
  FC:1μg/Test
  IF:1:100-500

•  

  

  Fig1: Sample:
  Brain(Rat) lysate at30ug;
  Lung(Rat) lysate at 30ug;
  Primary: Anti-Cyclin Eat 1:200;
  Secondary: HRP conjugated Goat-Anti-Rabbit IgG(bse-0295G) at 1: 3000;
  Predicted band size : 45kD
  Observed band size : 45kD

   

  

  Fig2: Tissue/cell: human laryngocarcinoma; 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-Cyclin-E Polyclonal Antibody, Unconjugated1:200, overnight at 4℃, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

   

  

  Fig3: Cell: NIH/3T3
  Concentration:1:100
  Host/Isotype:Rabbit/IgG
  Flow cytometric analysis of primary antibody  on NIH/3T3(green) compared with Rabbit IgG isotype control in the absence of primary antibody (blue) followed by Alexa Fluor 488-conjugated goat anti-rabbit IgG(H+L) secondary antibody .

   

  

  Fig4: Sample: Hela Lysate at 40 ug
  Primary: Anti-Cyclin at 1/300 dilution
  Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/10000 dilution
  Predicted band size: 45 kD
  Observed band size: 48 kD

   

  

  Fig5: Sample:
  Brain(Mouse) Lysate at 40 ug
  Testis(Mouse) Lysate at 40 ug
  Primary: Anti-Cyclin E  at 1/300 dilution
  Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/10000 dilution
  Predicted band size: 45 kD
  Observed band size: 48 kD

   

  

  Fig6: Blank control (blue line): Mouse spleen cells (blue).
  Primary Antibody (green line): Rabbit Anti-Cyclin E1 antibody 
  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 70% ethanol (overninght at 4℃) 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.

   

  

  Fig7: Sample:
  MCF-7(Human) Cell Lysate at 30 ug
  Primary: Anti-Cyclin E1at 1/1000 dilution
  Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
  Predicted band size: 45 kD
  Observed band size: 48 kD

   

  

  Fig8: Sample:
  Placenta (Mouse) Lysate at 40 ug
  Primary: Anti-Cyclin E1 at 1/1000 dilution
  Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
  Predicted band size: 45 kD
  Observed band size: 48 kD

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