• Western blot analysis of extracts of Jurkat cells, using Phospho-ZAP70-Y493 antibody (STJ22449) at 1:1000 dilution. Jurkat cells were treated by Hydrogen Peroxide (2 mM) at 37 °C for 2 minutes after serum-starvation overnight. Jurkat cells were treated by Pervanadate (1 mM) at 37 °C for 30 minutes. Secondary antibody: HRP Goat Anti-rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% non-fat dry milk in TBST. Detection: ECL Basic Kit. Exposure time: 1s.
  • Immunofluorescence analysis of Jurkat cells using Phospho-ZAP70-Y493 Rabbit polyclonal antibody (STJ22449) at dilution of 1:100 (40x lens). Jurkat cells were treated by Hydrogen Peroxide (2 mM) at 37 °C for 2 minutes after serum-starvation overnight. Secondary antibody: Cy3 Goat Anti-Rabbit IgG (H+L) at 1:500 dilution. Blue: DAPI for nuclear staining.
  • Immunohistochemistry of paraffin-embedded human breast carcinoma tissue, using Phospho-ZAP70-Y493 antibody (STJ22449).
  • Immunohistochemistry analysis of Phospho-ZAP70-Y493 in paraffin-embedded human breast carcinoma tissue, using Phospho-ZAP70-Y493 Rabbit polyclonal antibody (STJ22449). Perform microwave antigen retrieval with 10 mM Tris/EDTA buffer pH 9. 0 before commencing with immunohistochemistry staining protocol.
  • Immunofluorescence analysis of Jurkat cells using Phospho-ZAP70-Y493 rabbit polyclonal antibody (STJ22449) at dilution of 1:100 (40x lens). Jurkat cells were treated by Hydrogen Peroxide (2 mM) at 37 °C for 2 minutes after serum-starvation overnight. Blue: DAPI for nuclear staining.
  • Western blot analysis of lysates from Jurkat cells, using Phospho-ZAP70-Y493 Rabbit polyclonal antibody (STJ22449) at 1:1000 dilution. Jurkat cells were treated by Hydrogen Peroxide (2 mM) at 37 °C for 2 minutes after serum-starvation overnight. Jurkat cells were treated by Pervanadate (1 mM) at 37 °C for 30 minutes. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (STJS000856) at 1:10000 dilution. Lysates/proteins: 25 Mu g per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL Basic Kit. Exposure time: 1s.

Anti-Phospho-ZAP70-Y493 antibody (STJ22449)

SKU:
STJ22449

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Host: Rabbit
Applications: WB/IHC/IF
Reactivity: Human/Mouse/Rat
Note: STRICTLY FOR FURTHER SCIENTIFIC RESEARCH USE ONLY (RUO). MUST NOT TO BE USED IN DIAGNOSTIC OR THERAPEUTIC APPLICATIONS.
Short Description: Rabbit polyclonal antibody anti-Phospho-ZAP70-Y493 is suitable for use in Western Blot, Immunohistochemistry and Immunofluorescence research applications.
Clonality: Polyclonal
Conjugation: Unconjugated
Isotype: IgG
Formulation: PBS with 0.01% Thimerosal, 50% Glycerol, pH7.3.
Purification: Affinity purification
Dilution Range: WB 1:500-1:1000
IHC-P 1:50-1:100
IF/ICC 1:50-1:200
Storage Instruction: Store at-20°C for up to 1 year from the date of receipt, and avoid repeat freeze-thaw cycles.
Gene Symbol: ZAP70
Gene ID: 7535
Uniprot ID: ZAP70_HUMAN
Immunogen: A synthetic phosphorylated peptide around Y493 of human ZAP70 (NP_001070.2).
Immunogen Sequence: SYYTA
Tissue Specificity Expressed in T- and natural killer cells. Also present in early thymocytes and pro/pre B-cells.
Post Translational Modifications 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.
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).
Protein Name Tyrosine-Protein Kinase Zap-70
70 Kda Zeta-Chain Associated Protein
Syk-Related Tyrosine Kinase
Database Links Reactome: R-HSA-202430
Reactome: R-HSA-202433
Reactome: R-HSA-9013407
Cellular Localisation Cytoplasm
Cell Membrane
Peripheral Membrane Protein
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
Alternative Antibody Names Anti-Tyrosine-Protein Kinase Zap-70 antibody
Anti-70 Kda Zeta-Chain Associated Protein antibody
Anti-Syk-Related Tyrosine Kinase antibody
Anti-ZAP70 antibody
Anti-SRK antibody

Information sourced from Uniprot.org

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