Mouse anti-ZAP70 Monoclonal Antibody(1475CT217.34.65)

Mouse anti-ZAP70 Monoclonal Antibody(1475CT217.34.65)描述别名宿主特异性反应种属应用分子量类型克隆号同种型储存/保存方法研究领域背景说明细胞定位UniProt参考文献

概述
描述
Purified Mouse Monoclonal Antibody (Mab)
别名
ZAP70抗体;Tyrosine-protein kinase ZAP-70; 70 kDa zeta-chain associated protein; Syk-related tyrosine kinase; ZAP70; SRK
宿主
Mouse
特异性
This ZAP70 antibody is generated from a mouse immunized with a recombinant protein.
反应种属
Human
应用
IF~~1:25
FC~~1:25
WB~~1:4000
分子量
Predicted molecular weight: 70kD
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
性能
类型
Monoclonal Antibody
克隆号
1475CT217.34.65
同种型
IgG1,k
储存/保存方法
Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
研究领域
Crown Antibodies
靶标
背景说明
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. Subsequently, ZAP70 phosphorylates at least 2 essential adapter proteins: LAT and LCP2. 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. During thymocyte development, ZAP70 promotes survival and cell-cycle progression of developing thymocytes before positive selection (when cells are still CD4/CD8 double negative). Additionally, ZAP70-dependent signaling pathway may also contribute to primary B-cells formation and activation through B-cell receptor (BCR).
细胞定位
Cytoplasm. Cell membrane; Peripheral membrane protein. Note=In quiescent T-lymphocytes, it is cytoplasmic. Upon TCR activation, it is recruited at the plasma membrane by interacting with CD247/CD3Z. Colocalizes together with RHOH in the immunological synapse. RHOH is required for its proper localization to the cell membrane and cytoskeleton fractions in the thymocytes (By similarity).
UniProt
P43403
参考文献
参考文献
Chan A.C.,et al.Cell 71:649-662(1992).
Kuroyama H.,et al.Biochem. Biophys. Res. Commun. 315:935-941(2004).
Hillier L.W.,et al.Nature 434:724-731(2005).
Arpaia E.,et al.Cell 76:947-958(1994).
Isakov N.,et al.J. Exp. Med. 181:375-380(1995).

实验结果图

Mouse anti-ZAP70 Monoclonal Antibody(1475CT217.34.65)

Overlay histogram showing Jurkat cells stained with JP100494 (green line). The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 90% methanol for 10 min. The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (JP100494, 1:25 dilution) for 60 min at 37ºC. The secondary antibody used was Goat-Anti-Mouse IgG, DyLight® 488 Conjugated Highly Cross-Adsorbed(NA168821)) at 1/400 dilution for 40 min at 37ºC. Isotype control antibody (blue line) was mouse IgG1 (1μg/1×10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.

Mouse anti-ZAP70 Monoclonal Antibody(1475CT217.34.65)

All lanes : Anti-ZAP70 Antibody at 1:4000 dilution Lane 1: Jurkat whole cell lysates Lane 2: MOLT-4 whole cell lysates Lysates/proteins at 20 μg per lane. Secondary Goat Anti-mouse IgG, (H+L), Peroxidase conjugated at 1/10000 dilution Predicted band size : 70 kDa Blocking/Dilution buffer: 5% NFDM/TBST.