zeta-Chain (TCR) Associated Protein Kinase 70kDa (ZAP70) antibody

Antigen: zeta-Chain (TCR) Associated Protein Kinase 70kDa (ZAP70)

Synonyms: SRK, STD, TZK, ZAP-70, FLJ17670, FLJ17679, Srk, mur, 70kDa, mrtle, AI327364, MGC108902, zap70, MGC80670, MGC88962, ZAP70

Clonality: Polyclonal

Host: Rabbit

Reactivity: Human

Application: Western Blotting (WB)

Catalog no.: ABIN94502

Additional Information

Alternative name: ZAP70

Gene ID: 2q12

Immunogen: Bacterially expressed fusion protein representing C-terminal part (160 amino acids) ofhuman ZAP70 with histidine tag

Format: Purified

Description: The ZAP70 (zeta-associated protein of 70 kDa) tyrosine kinase was identified as a tyrosinephosphoprotein that associates with TCR zeta subunit and undergoes tyrosinephosphorylation following TCR stimulation. ZAP70 is a Syk family tyrosine kinase primarilyexpressed in T and NK cells that plays an essential role in signaling through the TCR.TCR-mediated activation of T cells is crucial to the immune response. In humans, ZAP70gene mutations resulting in lower ZAP70 protein expression levels or expression ofcatalytically inactive ZAP70 proteins, have been identified. ZAP70 deficiency results in theabsence of mature CD8+ T cells and the prevention of TCR-mediated activation of CD4+ Tcells, and it can lead to severe combined immunodeficiency. ZAP70 is cytosolic proteinmigrating at 70 kDa in SDS-PAGE. It contains two N-terminal SH2 domains (Src homologydomain 2) and a C-terminal kinase domain. Crystal structure of the ZAP70 SH2 domains incomplex with a TCR zeta subunit peptide was described. During T cell activation, thebinding of ZAP70 SH2 domains to the phosphorylated zeta subunit on the activated TCRcomplex causes a colocalization with the Lck tyrosine kinase that phosphorylates ZAP70 onTyr493 in the activation loop. ZAP70 autophosphorylates multiple tyrosines in the regionbetween the SH2 domains and the kinase domain, including the binding sites for additionalSH2-containing signaling proteins such as SLP76, LAT, Lck, PLCgamma1, Vav, Shc,Ras-GAP, and Abl. ZAP70-mediated activation of these downstream effectors leads to therelease of intracellular calcium stores, and the transcription of interleukin-2 and other genesimportant for an immune response.

Specificity: The polyclonal antibody recognizes C-terminal part of human ZAP70 protein tyrosine kinase.ZAP70 is a molecule susceptible to degradation. It is recommended to use freshlyprepared cell lysates (protease inhibitors are essential) to avoid non-specific staining ofdegradation products.

Application Details

Application Notes: Western Blotting - Recommended dilution: 1 µg/ml. Positive control: JURKAT T cell leukemia cell line. Sample preparation: Resuspend approx. 50 mil. cells in 1 ml cold Lysis buffer (1%laurylmaltoside in 20 mM Tris/Cl, 100 mM NaCl pH 8. 2, 50 mM NaF including Proteaseinhibitor Cocktail). Incubate 60 min on ice. Centrifuge to remove cell debris. Mix lysate withreducing Laemmli SDS-PAGE sample buffer. Application note: Reducing conditions.

Concentration: 1 mg/ml

Purity: > 95% (by SDS-PAGE)

Purification: Purified from rabbit serum by protein-A afinity chromatography

Buffer: Phosphate buffered saline (PBS) with 15 mM sodium azide, approx. pH 7.4

Storage: Store at 2-8 o C. Do not use after expiration date stamped on vial label. For long-term storage aliquot and store at -20 o C. Avoid freeze/thaw cycles.

Research Area: Signaling, Tags/Labels, Immunology, Cancer, CD Antigens, Surface Receptors of Immune Cells

Restrictions: For Research Use only