Anti-Zap70 Antibody

Product Name

Anti-Zap70 Antibody

View all ZAP70 Antibodies

SKU/Catalog Number

M00754-2

Size

80 µl

Description

Boster Bio Anti-Zap70 Antibody (Catalog # M00754-2). Tested in WB, Flow Cytometry, IF application(s). This antibody reacts with Human, Mouse.

Storage & Handling

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.

Cite This Product

Anti-Zap70 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00754-2)

Host

Mouse

Contents

Purified monoclonal antibody supplied in PBS with 0.09% (W/V) sodium azide.

Clonality

Monoclonal

Clone Number

1484CT290.68.62

Isotype

IgG2a,k

Immunogen

This Zap70 antibody is generated from a mouse immunized with a recombinant protein.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Reactive Species

M00754-2 is reactive to ZAP70 in Human, Mouse

Applications

M00754-2 is guaranteed for Flow Cytometry, IF, WB Boster Guarantee

Observed Molecular Weight

Calculated molecular weight

69.872kDa

Background of ZAP70

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).

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

IF: 1:25
WB: 1:2000
FC: 1:25

Validation Images & Assay Conditions

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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervical epithelial adenocarcinoma cell line) cells labeling Zap70 with M00754-2 at 1/25 dilution, followed by Dylight® 488-conjugated goat anti-mouse IgG secondary antibody at 1/200 dilution (green). Immunofluorescence image showing cytoplasm staining on HeLa cell line. Cytoplasmic actin is detected with Dylight® 554 Phalloidin at 1/100 dilution (red).The nuclear counter stain is DAPI (blue).

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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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Overlay histogram showing Jurkat cells stained with M00754-2 (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 (M00754-2, 1:25 dilution) for 60 min at 37ºC. The secondary antibody used was Goat-Anti-Mouse IgG, DyLight® 488 Conjugated Highly Cross-Adsorbed at 1/400 dilution for 40 min at 37ºC. Isotype control antibody (blue line) was mouse IgG2a (1g/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.

Gene/Protein Information For ZAP70 (Source: Uniprot.org, NCBI)

Alternative Names

70 kDa zeta-associated protein; EC 2.7.10; EC 2.7.10.2; mrtle; mur; SRK; SRKFLJ17679; STD; STDFLJ17670; Syk-related tyrosine kinase; tyrosine-protein kinase ZAP-70; TZK; ZAP70; ZAP-70; zeta-chain (TCR) associated protein kinase (70 kD); zeta-chain (TCR) associated protein kinase 70kDa; zeta-chain associated protein kinase, 70kD ZAP70 ADMIO2, IMD48, SRK, STCD, STD, TZK, ZAP-70 zeta chain of T cell receptor associated protein kinase 70 tyrosine-protein kinase ZAP-70|70 kDa zeta-associated protein|70 kDa zeta-chain associated protein|syk-related tyrosine kinase|zeta chain of T cell receptor associated protein kinase 70kDa|zeta-chain (TCR) associated protein kinase 70kDa|zeta-chain associated protein kinase, 70kD

For more info on ZAP70, check out the ZAP70 Infographic

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In this infographic, you will see the following information for ZAP70: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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