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>   首页   >   产品   >   一抗   >   信号转导   >   Zap70 Antibody   

Zap70 Antibody

Purified Mouse Monoclonal Antibody (Mab)

     
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  • 3 - Zap70 Antibody AM8462b
    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0. 1% Triton X-100 permeabilized HeLa (human cervical epithelial adenocarcinoma cell line) cells labeling Pdx1 with AM8462b at 1/25 dilution, followed by Dylight® 488-conjugated goat anti-mouse IgG (NA166821) secondary antibody at 1/200 dilution (green). Immunofluorescence image showing cytoplasm staining on HeLa cell line. Cytoplasmic actin is detected with Dylight® 554 Phalloidin (PD18466410) at 1/100 dilution (red). The nuclear counter stain is DAPI (blue).
  • 4 - Zap70 Antibody AM8462b
    Overlay histogram showing Jurkat cells stained with AM8462b (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 (AM8462b, 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 IgG2a (1μg/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.
  • 1 - Zap70 Antibody AM8462b
    All lanes : Anti-Zap70 Antibody at 1:2000 dilution Lane 1: Jurkat whole cell lysates Lane 2: MOLT-4 whole cell lysates Lane 3: mouse thymus 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.
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Product info
Application
  • Applications Legend:
  • E=ELISA
  • WB=Western Blotting
  • IHC=Immunohistochemistry
  • IHC-P=Immunohistochemistry (Paraffin)
  • IP=Immunoprecipitation
  • IF=Immunofluorescence
  • IC=Immunochemistry
  • ICC=Immunocytochemistry
  • FC=Flow Cytometry
  • DB=Dot Blot
IF, FC, WB, E
Primary Accession P43404
Reactivity Human, Mouse
Host Mouse
Clonality monoclonal
Isotype IgG2a,k
Clone Names 1484CT290.68.62
Calculated MW 70112 Da
Additional info
Gene ID 22637
Other Names Tyrosine-protein kinase ZAP-70, 70 kDa zeta-chain associated protein, Syk-related tyrosine kinase, Zap70, Srk, Zap-70
Target/Specificity This Zap70 antibody is generated from a mouse immunized with a recombinant protein.
Dilution IF~~1:25
FC~~1:25
WB~~1:2000
StorageMaintain 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.
PrecautionsZap70 Antibody is for research use only and not for use in diagnostic or therapeutic procedures.
Protein Information
Name Zap70
Synonyms Srk, Zap-70
Function 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 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-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).
Cellular Location Cytoplasm. Cell membrane; Peripheral membrane protein. Note=In quiescent T-lymphocytes, ZAP70 is cytoplasmic. Upon TCR activation, it is recruited at the plasma membrane by interacting with 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
Tissue Location Isoform 1 and isoform 2 are expressed in thymus, spleen and lymph nodes.
Research Areas

BACKGROUND

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 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-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).

REFERENCES

Gauen L.K.T.,et al.Mol. Cell. Biol. 14:3729-3741(1994).
Wiest D.L.,et al.Immunity 6:663-671(1997).
Kuroyama H.,et al.Biochem. Biophys. Res. Commun. 315:935-941(2004).
Ikeda T.,et al.Submitted (APR-2002) to the EMBL/GenBank/DDBJ databases.
Carninci P.,et al.Science 309:1559-1563(2005).

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