Phospho-ZAP70/Syk (Tyr319, Tyr352) Monoclonal Antibody (n3kobu5), PE-Cyanine7

货号：25-9006-41,25-9006-42

规格：25 Tests,100 Tests

价格：1406,3294

产品类型：流式抗体

品牌：eBioscience

物种：人/小鼠

宿主：小鼠

抗体亚型：IgG2b, kappa

克隆号：n3kobu5

荧光染料：PE-Cy7

类型:	单抗	同型对照：	Mouse IgG2b kappa Isotype Control (eBMG2b), PE-Cyanine7
浓度：	5 µL/Test	用法：	5 µL (0.06 µg)/test(Flow)

产品详细信息Description: This n3kobu5 monoclonal antibody recognizes human and mouse zeta chain-associated protein of 70 kD (also known as ZAP-70) and spleen tyrosine kinase (also known as SYK) when phosphorylated on Y319 and Y352, respectively. ZAP-70 and SYK are members of the SYK protein tyrosine kinase (PTK) family that are phosphorylated and activated by Src family PTKs. ZAP-70/SYK Y319/Y352 are located in the so-called interdomain of ZAP-70/SYK (between the N-terminal dual SH2 domains and the C-terminal kinase domain).Phosphorylation of ZAP-70 Y319 by Lck is necessary for T cell receptor (TCR)-dependent association of ZAP-70 with Lck and phospholipase C gamma and subsequent activation of calcium-dependent and Ras signaling cascades. SYK Y352 phosphorylation by Fyn/Lyn is critical for propagation of B cell receptor (BCR) signaling and for B cell development.Specificity of this n3kobu5 clone was determined by ELISA, flow cytometry, and western blotting.Applications Reported: This n3kobu5 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.Applications Tested: This n3kobu5 antibody has been pre-diluted and tested by intracellular staining and flow cytometric analysis of sodium pervanadate-treated mouse splenocytes using the Intracellular Fixation and Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Please refer to "Staining Intracellular Antigens for Flow Cytometry, Protocol A: Two step protocol for intracellular (cytoplasmic) proteins" located atwww.thermofisher.com/flowprotocols. This may be used at 5 µL (0.06 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222-49) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-57) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.Excitation: 488-561 nm; Emission: 775 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser靶标信息ZAP70, a 70 kDa member of the Syk tyrosine kinase family, plays a central role in lymphocyte activation and development, and is implicated in several immune disorders. Upon T cell antigen receptor (TCR) engagement, ZAP70 is phosphorylated on tyrosines 292, 315 and 319 in the interdomain B, located between the SH2 and kinase domains. Phosphorylation of both tyrosines 315 (a Vav binding site) and 319 (a Lck binding site) enhances ZAP70 function in mediating lymphocyte signaling, while tyrosine 292terminates the transient activation of ZAP70 and attentuates lymphocyte signaling.

数据

Phospho-ZAP70/Syk (Tyr319, Tyr352) Antibody (25-9006-42) in FlowC57BL/6 mouse splenocytes were untreated (orange histogram) or treated for 5 minutes with hydrogen peroxide-activated sodium pervanadate. Cells were then stained intracellularly, using the Intracellular Fixation and Permeabilization Buffer Set (Product # 88-8824-00) and protocol, with CD3 Monoclonal Antibody, eFluor 450 (Product # 48-0032-82) and 0.06 µg of Phospho-ZAP70/Syk (Tyr319, Tyr352) Monoclonal Antibody, PE-Cyanine7 (purple histogram). Lymphocytes in the CD3-positive gate were used for analysis.

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