ARG66758

anti-ZAP70 antibody [SQab20191]

anti-ZAP70 antibody [SQab20191] for IHC-Formalin-fixed paraffin-embedded sections,Western blot and Human

nuts_pic
1

Overview

Product Description Recombinant Rabbit Monoclonal antibody [SQab20191] recognizes ZAP70
Tested Reactivity Hu
Tested Application IHC-P, WB
Host Rabbit
Clonality Monoclonal
Clone SQab20191
Isotype IgG
Target Name ZAP70
Antigen Species Human
Immunogen Synthetic peptide within aa. 200-350 of Human ZAP70.
Conjugation Un-conjugated
Alternate Names STD; SRK; STCD; 70 kDa zeta-chain associated protein; Tyrosine-protein kinase ZAP-70; TZK; Syk-related tyrosine kinase; ZAP-70; EC 2.7.10.2

Application Instructions

Application Suggestion
Tested Application Dilution
IHC-P1:100 - 1:200
WB1:1000
Application Note IHC-P: Antigen Retrieval: Heat mediation was performed in Tris/EDTA buffer (pH 9.0).
* The dilutions indicate recommended starting dilutions and the optimal dilutions or concentrations should be determined by the scientist.
Positive Control WB: Jurkat
IHC-P: Tonsil tissue
Observed Size ~ 70 kDa

Properties

Form Liquid
Purification Purification with Protein A.
Buffer PBS, 0.01% Sodium azide, 40% Glycerol and 0.05% BSA.
Preservative 0.01% Sodium azide
Stabilizer 40% Glycerol and 0.05% BSA
Storage Instruction For continuous use, store undiluted antibody at 2-8°C for up to a week. For long-term storage, aliquot and store at -20°C. Storage in frost free freezers is not recommended. Avoid repeated freeze/thaw cycles. Suggest spin the vial prior to opening. The antibody solution should be gently mixed before use.
Note For laboratory research only, not for drug, diagnostic or other use.

Bioinformation

Database Links

GeneID: 7535 Human ZAP70

Swiss-port # P43403 Human Tyrosine-protein kinase ZAP-70

Gene Symbol ZAP70
Gene Full Name zeta-chain (TCR) associated protein kinase 70kDa
Background This gene encodes an enzyme belonging to the protein tyrosine kinase family, and it plays a role in T-cell development and lymphocyte activation. This enzyme, which is phosphorylated on tyrosine residues upon T-cell antigen receptor (TCR) stimulation, functions in the initial step of TCR-mediated signal transduction in combination with the Src family kinases, Lck and Fyn. This enzyme is also essential for thymocyte development. Mutations in this gene cause selective T-cell defect, a severe combined immunodeficiency disease characterized by a selective absence of CD8-positive T-cells. Two transcript variants that encode different isoforms have been found for this gene. [provided by RefSeq, Jul 2008]
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. 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-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. 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). [UniProt]
Cellular Localization Cytoplasm. Cell membrane; Peripheral membrane protein. Note=In quiescent T-lymphocytes, it is cytoplasmic. Upon TCR activation, it is recruited at the plasma membrane by interacting with CD247/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 (By similarity). [UniProt]
Calculated MW 70 kDa
PTM 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. [UniProt]

Customer's Feedback

nuts_pic      Excellent

Review for anti-ZAP70 antibody [SQab20191]

Application:WB

Sample:Jurkat

Sample Loading Amount:10 µg

Primary Antibody Dilution Factor:1:1000

Primary Antibody Incubation Time:overnight

Primary Antibody Incubation Temperature:4 ºC