ZAP-70/Syk (pTyr319) antibody (Alexa Fluor 488)

Catalog No. ABIN1176923

The Mouse Monoclonal anti- antibody has been validated for ICS. It is suitable to detect in samples from Human, Mouse and Rat. There are 3+ publications available.

Quick Overview for ZAP-70/Syk (pTyr319) antibody (Alexa Fluor 488) (ABIN1176923)

Target: ZAP-70/Syk

Reactivity: Human, Mouse, Rat

Host: Mouse

Clonality: Monoclonal

Conjugate: Alexa Fluor 488

Application: Intracellular Staining (ICS)

Clone: 17A-P

Product Details

Binding Specificity: pTyr319

Brand: BD Phosflow™

Purification: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Immunogen: Human phosphorylated ZAP70 Peptide

Isotype: IgG1

Application Details

Application Notes: Jurkat cells treated with H2O2 are suggested as a positive control. However, other cell types or methods may also be used for detection of phosphorylated ZAP70.

Sample Volume: 20 μL

Restrictions: For Research Use only

Handling

Format: Liquid

Buffer: Aqueous buffered solution containing BSA and ≤0.09 % sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: 4 °C

Storage Comment: Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The antibody was conjugated to Alexa Fluor® 488 under optimum conditions, and unreacted Alexa Fluor® 488 was removed.

References

Di Bartolo, Mège, Germain, Pelosi, Dufour, Michel, Magistrelli, Isacchi, Acuto: "Tyrosine 319, a newly identified phosphorylation site of ZAP-70, plays a critical role in T cell antigen receptor signaling." in: The Journal of biological chemistry, Vol. 274, Issue 10, pp. 6285-94, (1999) (PubMed).

Arpaia, Shahar, Dadi, Cohen, Roifman: "Defective T cell receptor signaling and CD8+ thymic selection in humans lacking zap-70 kinase." in: Cell, Vol. 76, Issue 5, pp. 947-58, (1994) (PubMed).

Chan, Kadlecek, Elder, Filipovich, Kuo, Iwashima, Parslow, Weiss: "ZAP-70 deficiency in an autosomal recessive form of severe combined immunodeficiency." in: Science (New York, N.Y.), Vol. 264, Issue 5165, pp. 1599-601, (1994) (PubMed).

Target Details

Target: ZAP-70/Syk

Alternative Name: Zap70 ,Syk

Background: ZAP70 is a protein tyrosine kinase (PTK) that associates with the z subunit of the T cell antigen receptor (TCR) and undergoes tyrosine phosphorylation following TCR stimulation. ZAP70 contains two SH2-like domains with the PTK domain located at the C-terminus. It appears that both ZAP70 and Syk are recruited to the phosphorylated CD3 and z subunits after TCR stimulation. TCR stimulation leads to autophosphorylation of ZAP70 at Tyr-315 amd Tyr-319, and mutation of the Tyr-319 site dramatically impairs TCR signaling. In addition, TCR-mediated Lck activity leads to phosphorylation of ZAP70 on Tyr-493 in the regulatory loop of the kinase domain leading to upregulation of ZAP70 kinase activity. The significance of ZAP70 activation in mediating TCR signal transduction has been confirmed by showing that ZAP70 activity is absent in an autosomal recessive form of severe combined immunodeficiency (SCID). This is due to mutations affecting the ZAP70 kinase domain which affect the stability of the protein and TCR signaling. Clone 17A/P-ZAP70 recognizes the phosphorylated form of ZAP70 (Y319). It also cross-reacts with SYK (Y352) due to homology of the phosphorylation site with ZAP70 (Y319). The PE-conjugated format has been evaluated using human and mouse model systems. The unconjugated form of the antibody (Cat. No. 612574) has also been shown to work in western blot analysis on human, mouse, and rat cells.
Synonyms: ZAP-70, SRK, STD, TZK, Zeta-chain associated protein kinase, 70kD