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Invitrogen
Phospho-ZAP70/Syk (Tyr319, Tyr352) Monoclonal Antibody (n3kobu5), PE, eBioscience™
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Phospho-ZAP70/Syk (Tyr319, Tyr352) Antibody (12-9006-42) in Flow
Intracellular staining of untreated (orange histogram) or 5-minute hydrogen peroxide-activated sodium pervanadate-treated (purple histogram) mouse spleen cells with Anti-Human/Mouse phospho-ZAP-70/SYK (Y319/Y352) PE using the Intracellular Fixation & Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Lymphocytes in the CD3+ (left) or B220+ (right) gates were used for analysis.
产品信息
12-9006-42
应用
建议稀释比
已发表文章
产品规格
种属反应
Human,
Mouse
已发表种属
Mouse
宿主/亚型
Mouse
/ IgG2b, kappa
分类
Monoclonal
类型
Antibody
克隆号
n3kobu5
偶联物
激发/发射光谱
565/576 nm
查看光谱
形式
Liquid
浓度
5 µL/Test
纯化类型
Affinity chromatography
保存液
PBS, pH 7.2, with 0.2% BSA
内含物
0.09% sodium azide
保存条件
4° C, store in dark, DO NOT FREEZE!
运输条件
Ambient (domestic); Wet ice (international)
RRID
AB_2572665
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-titrated and tested by intracellular staining followed by flow cytometric analysis of stimulated normal human peripheral blood cells. This can be used at 5 µL (0.03 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.
Staining Protocol: All protocols work well for this monoclonal antibody. Use of Protocol A: Two-step protocol: intracellular (cytoplasmic) proteins allows for the greatest flexibility for detection of surface and intracellular (cytoplasmic) proteins. Use of Protocol B: One-step protocol: intracellular (nuclear) proteins is recommended for staining of transcription factors in conjunction with surface and phosphorylated intracellular (cytoplasmic) proteins. Protocol C: Two-step protocol: Fixation/Methanol allows for the greatest discrimination of phospho-specific signaling between unstimulated and stimulated samples, but with limitations on the ability to stain specific surface proteins (refer to "Clone Performance Following Fixation/Permeabilization" located in the Best Protocols Section under the Resources tab online). All Protocols can be found in the Flow Cytometry Protocols: "Staining Intracellular Antigens for Flow Cytometry Protocol" located in the Best Protocols Section under the Resources tab online.
Excitation: 488-561 nm; Emission: 578 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.
Filtration: 0.2 µm post-manufacturing filtered.
靶标信息
ZAP-70 is a cytosolic protein tyrosine kinase that is a member of the Syk family of proteins. ZAP-70 is expressed exclusively in T-cells and natural killer cells and is required for T-cell receptor activation. 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 292 terminates the transient activation of ZAP70 and attentuates lymphocyte signaling.
仅用于科研。不用于诊断过程。未经明确授权不得转售。
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生物信息学
蛋白别名: 70 kDa zeta-associated protein; 70 kDa zeta-chain associated protein; EC 2.7.10.2; FLJ17670; FLJ17679; kinase ZAP70; Selective T cell defect; Syk-related tyrosine kinase; Tyrosine-protein kinase ZAP-70; Tyrosine-protein kinase ZAP-70;TZK; ZA70; Zeta chain associated protein kinase 70kDa; zeta chain of T cell receptor associated protein kinase 70kDa; zeta-chain (TCR) associated protein kinase 70kDa; zeta-chain associated protein kinase, 70kD
基因别名: ADMIO2; IMD48; mrtle; mur; SRK; STCD; STD; TZK; ZAP-70; ZAP70
UniProt ID: (Human) P43403, (Mouse) P43404
Entrez Gene ID: (Human) 7535, (Mouse) 22637
phosphotyrosine binding
protein tyrosine kinase activity
non-membrane spanning protein tyrosine kinase activity
receptor binding
protein binding
ATP binding
nucleotide binding
protein kinase activity
kinase activity
transferase activity
transferase activity, transferring phosphorus-containing groups
adaptive immune response
macrophage activation involved in immune response
neutrophil activation involved in immune response
protein phosphorylation
inflammatory response
immune response
transmembrane receptor protein tyrosine kinase signaling pathway
peptidyl-tyrosine phosphorylation
T cell differentiation
positive regulation of cell adhesion mediated by integrin
intracellular signal transduction
peptidyl-tyrosine autophosphorylation
T cell activation
B cell activation
positive regulation of mast cell degranulation
beta selection
positive thymic T cell selection
negative thymic T cell selection
innate immune response
positive regulation of B cell differentiation
positive regulation of T cell differentiation
positive regulation of alpha-beta T cell differentiation
positive regulation of alpha-beta T cell proliferation
positive regulation of calcium-mediated signaling
T cell receptor signaling pathway
B cell receptor signaling pathway
T cell aggregation
T cell migration
regulation of platelet aggregation
immune system process
phosphorylation
cell migration
cell differentiation
thymic T cell selection
protein autophosphorylation
