ZAP70 Mouse Monoclonal Antibody [A1B7]
Catalog# EM1902-28
ZAP70 Mouse Monoclonal Antibody [A1B7]
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WB
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IF-Cell
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FC
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
ZAP70 Mouse Monoclonal Antibody [A1B7]
Antibody Type
Mouse Monoclonal Antibody
Immunogen
Recombinant protein within human ZAP70 aa 200-500.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, FC
Molecular Weight
Predicted band size: 70 kDa
Positive Control
Jurkat cell lysate, mouse spleen tissue lysate, mouse thymus tissue lysate, rat spleen tissue lysate, rat thymus tissue lysate, LOVO, Jurkat.
Conjugation
unconjugated
Clone Number
A1B7
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage Buffer
1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG2a
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:500-1:2,000
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IF-Cell
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1:50
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FC
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1:50-1:100
Target
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 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.
Background References
1. Hu H. et. al. Otud7b facilitates T cell activation and inflammatory responses by regulating Zap70 ubiquitination. J. Exp. Med. 213:399-414(2016)
Sequence Similarity
Belongs to the protein kinase superfamily. Tyr protein kinase family. SYK/ZAP-70 subfamily.
Tissue Specificity
Expressed in T- and natural killer cells. Also present in early thymocytes and pro/pre B-cells.
Post-translational Modification
Phosphorylated on tyrosine residues upon T-cell antigen receptor (TCR) stimulation. Phosphorylation of Tyr-315 and Tyr-319 are essential for ZAP70 positive function on T-lymphocyte activation whereas Tyr-292 has a negative regulatory role. Within the C-terminal kinase domain, Tyr-492 and Tyr-493 are phosphorylated after TCR induction, Tyr-492 playing a negative regulatory role and Tyr-493 a positive. Tyr-493 is dephosphorylated by PTN22.; Ubiquitinated in response to T cell activation. Deubiquitinated by OTUD7B.
Subcellular Location
Cell membrane, cytoplasm.
Synonyms
70 kDa zeta associated protein antibody
70 kDa zeta-associated protein antibody
EC 2.7.10.2 antibody
FLJ17670 antibody
FLJ17679 antibody
Selective T cell defect antibody
SRK antibody
STD antibody
Syk related tyrosine kinase antibody
Syk-related tyrosine kinase antibody
ExpandImages
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☑ Relative expression (RE)
Western blot analysis of ZAP70 on different lysates with Mouse anti-ZAP70 antibody (EM1902-28) at 1/1,000 dilution.
Lane 1: Jurkat cell lysate
Lane 2: Ramos cell lysate (negative)
Lane 3: Mouse spleen tissue lysate
Lane 4: Mouse thymus tissue lysate
Lane 5: Rat spleen tissue lysate
Lane 6: Rat thymus tissue lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 70 kDa
Observed band size: 70 kDa
Exposure time: 1 minute 21 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1902-28) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature. -
ICC staining of ZAP70 in LOVO cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (EM1902-28, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Mouse IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
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Flow cytometric analysis of ZAP70 was done on Jurkat cells. The cells were fixed, permeabilized and stained with the primary antibody (EM1902-28, 1/100) (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 1/500 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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