Anti-ZAP70 antibody [A1B5]

Anti-ZAP70 antibody [A1B5]

• 概述

• 产品描述Tyrosine kinase that plays an essential role in regulation of the adaptive immune response. Regulates motility, adhesion and cytokine expression of mature T-cells, as well as thymocyte development. Contributes also to the development and activation of primary B-lymphocytes. When antigen presenting cells (APC) activate T-cell receptor (TCR), a serie of phosphorylations lead to the recruitment of ZAP70 to the doubly phosphorylated TCR component CD247/CD3Z through ITAM motif at the plasma membrane. This recruitment serves to localization to the stimulated TCR and to relieve its autoinhibited conformation. Release of ZAP70 active conformation is further stabilized by phosphorylation mediated by LCK. In turn, a large number of signaling molecules are recruited and ultimately lead to lymphokine production, T-cell proliferation and differentiation. Furthermore, ZAP70 controls cytoskeleton modifications, adhesion and mobility of T-lymphocytes, thus ensuring correct delivery of effectors to the APC. ZAP70 is also required for TCR-CD247/CD3Z internalization and degradation through interaction with the E3 ubiquitin-protein ligase CBL and adapter proteins SLA and SLA2. Thus, ZAP70 regulates both T-cell activation switch on and switch off by modulating TCR expression at the T-cell surface.
• 产品名称Anti-ZAP70 antibody [A1B5]
• 分子量70 kDa
• 种属反应性Human
• 验证应用WB,IHC-P,FC,ICC
• 抗体类型小鼠单抗
• 免疫原Recombinant protein within human ZAP70 aa 250-480.
• 偶联Non-conjugated

• 性能

• 形态Liquid
• 浓度2 mg/mL
• 存放说明Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
• 存储缓冲液1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
• 亚型IgG2a
• 纯化方式Protein G purified.
• 亚细胞定位Cell membrane, cytoplasm.
• 其它名称
  • 70 kDa zeta associated protein antibody
  • 70 kDa zeta-associated protein antibody
  • EC 2.7.10.2 antibody
  more

• 应用

  WB: 1:500-1:2,000
  IHC-P: 1:50-1:100
  FC: 1:50-1:100
  ICC: 1:50

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  Fig1: Western blot analysis of ZAP70 on Jurkat cell lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.

   

  

  Fig2: Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-ZAP70 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

   

  

  Fig3: Flow cytometric analysis of ZAP70 was done on Jurakt cells. The cells were fixed, permeabilized and stained with the primary antibody  (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Mouse IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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