Mouse Monoclonal ZAP70 Antibody

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     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.    

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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 P32984 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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     Overlay histogram showing Jurkat cells stained with P32984 (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 (P32984,  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/1x10^6 cells) used under the same conditions.  Acquisition of >10, 000 events was performed.    

ZAP-70 (zeta-chain-associated protein kinase 70) is a cytoplasmic protein tyrosine kinase critical for T-cell receptor (TCR) signaling and immune system regulation. It belongs to the Syk kinase family and is primarily expressed in T cells and natural killer (NK) cells. Upon TCR activation, ZAP-70 is recruited to phosphorylated immunoreceptor tyrosine-based activation motifs (ITAMs) on the CD3ζ chain, where it initiates downstream signaling cascades essential for T-cell development, activation, and differentiation. Deficiencies in ZAP-70 function are linked to severe combined immunodeficiency (SCID), characterized by absent CD8+ T cells and dysfunctional CD4+ T cells.

ZAP-70 antibodies are laboratory tools used to detect and quantify ZAP-70 protein expression in research and diagnostics. In clinical settings, ZAP-70 expression is assessed in chronic lymphocytic leukemia (CLL) as a prognostic marker. High ZAP-70 levels correlate with unmutated immunoglobulin heavy-chain variable region (IGHV) genes, aggressive disease progression, and poorer outcomes, similar to CD38 expression. Detection methods include flow cytometry, immunohistochemistry (IHC), and Western blotting. Research applications focus on studying TCR signaling mechanisms, immune cell interactions, and therapeutic targeting. ZAP-70 inhibitors are also explored for modulating immune responses in autoimmune diseases or enhancing anti-tumor immunity. Its dual role as a signaling molecule and biomarker underscores its importance in immunology and hematology.