Invitrogen

Zap-70 Monoclonal Antibody (ZAP-70-6F7)

IP-mass spectrometry

4 篇参考文献

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Zap-70 Antibody in Immunocytochemistry (ICC/IF)

Zap-70 Antibody in Immunocytochemistry (ICC/IF)

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Zap-70 Antibody (13-0600) in ICC/IF

Immunofluorescence analysis of ZAP70 was performed using 70% confluent log phase Jurkat cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with ZAP70 (ZAP-70-6F7) Mouse Monoclonal Antibody (Product # 13-0600) at 2µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product ... View More

Zap-70 Antibody in Immunocytochemistry (ICC/IF)

Zap-70 Antibody in Western Blot (WB)

Zap-70 Antibody in Western Blot (WB)

Zap-70 Antibody

Zap-70 Antibody

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Zap-70 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of ZAP70 was performed using 70% confluent log phase Jurkat cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with ZAP70 (ZAP-70-6F7) Mouse Monoclonal Antibody (Product # 13-0600) at 2µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

Zap-70 Antibody in Western Blot (WB) — Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of Jurkat (Lane 1) and MOLT-4 (Lane 2). The blot was probed with Anti-ZAP70 Mouse Monoclonal Antibody (Product # 13-0600, 2 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.4 µg/mL, 1:2500 dilution). A 70 kDa band corresponding to ZAP70 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Zap-70 Antibody in Western Blot (WB) — Western blot analysis of Raji (lane 1) and Jurkat (lane 2) cells using Ms anti-ZAP-70 (Product # 13-0600)

Zap-70 Antibody — Antibody specificity was demonstrated by detection of differential basal expression of the target across cell models owing to their inherent genetic constitution. Relative expression of ZAP70 was observed in Raji and Jurkat cells in western blot using ZAP70 Monoclonal Antibody (Product # 13-0600). Raji (B-cell type) and Jurkat (T-cell type) cells are documented to have differential basal expression of ZAP70, where-in ZAP70 is expressed in T-cells and not in B-cells. {RE}

Zap-70 Antibody — IP-MS enrichment of ZAP70 (LFQ intensity): ZAP70 was enriched 68-fold from SR lysate compared to background proteins, using the optimized IP-MS workflow with Pierce MS-Compatible Magnetic IP Kit protein A/G (Product # 90409) and ZAP70 antibody (Product # 13-0600). The STRING database (www.string-db.org) was used to identify the protein interactor list. See more information on IP-MS verification of antibody selectivity. {IP-MS}

产品信息

13-0600

应用

建议稀释比

已发表文章

免疫印迹 (WB)

1-3 µg/mL

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免疫细胞化学 (ICC/IF)

2 µg/mL

-

酶联免疫吸附实验 (ELISA)

Assay-dependent

-

免疫沉淀 (IP)

Assay-dependent

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其他PubMed (Misc)

-

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产品规格

种属反应

Human, Mouse

已发表种属

Human, Mouse, Non-human primate

宿主/亚型

Mouse / IgG1, kappa

分类

Monoclonal

类型

Antibody

克隆号

ZAP-70-6F7

抗原

Peptide derived from a.a. 326-341 of human ZAP-70.

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偶联物

Unconjugated Unconjugated Unconjugated

形式

Liquid

浓度

0.5 mg/mL

纯化类型

Affinity chromatography

保存液

PBS, pH 7.4

内含物

0.1% sodium azide

保存条件

-20°C

运输条件

Ambient (domestic); Wet ice (international)

RRID

AB_2532997

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。

Product References

Lck dephosphorylation at Tyr-394 and inhibition of T cell antigen receptor signaling by Yersinia phosphatase YopH.

The Journal of biological chemistry

Alonso A,Bottini N,Bruckner S,Rahmouni S,Williams S,Schoenberger SP,Mustelin T

Fri Feb 06 00:00:00 EST 2004

Activation of ZAP-70 through specific dephosphorylation at the inhibitory Tyr-292 by the low molecular weight phosphotyrosine phosphatase (LMPTP).

The Journal of biological chemistry

Bottini N,Stefanini L,Williams S,Alonso A,Jascur T,Abraham RT,Couture C,Mustelin T

Fri Jul 05 00:00:00 EDT 2002

Activation of ZAP-70 through specific dephosphorylation at the inhibitory Tyr-292 by the low molecular weight phosphotyrosine phosphatase (LMPTP).

The Journal of biological chemistry

Bottini N,Stefanini L,Williams S,Alonso A,Jascur T,Abraham RT,Couture C,Mustelin T

Fri Jul 05 00:00:00 EDT 2002

Activation of ZAP-70 through specific dephosphorylation at the inhibitory Tyr-292 by the low molecular weight phosphotyrosine phosphatase (LMPTP).

The Journal of biological chemistry

Bottini N,Stefanini L,Williams S,Alonso A,Jascur T,Abraham RT,Couture C,Mustelin T

Fri Jul 05 00:00:00 EDT 2002

Activation of ZAP-70 through specific dephosphorylation at the inhibitory Tyr-292 by the low molecular weight phosphotyrosine phosphatase (LMPTP).

The Journal of biological chemistry

Bottini N,Stefanini L,Williams S,Alonso A,Jascur T,Abraham RT,Couture C,Mustelin T

Fri Jul 05 00:00:00 EDT 2002

Negative regulation of T-cell activation and autoimmunity by Mgat5 N-glycosylation.

Nature

Demetriou M,Granovsky M,Quaggin S,Dennis JW

13-0600 was used in immunoprecipitation to report that a deficiency in beta1,6 N-acetylglucosaminyltransferase V lowers T-cell activation thresholds by directly enhancing TCR clustering.

Thu Feb 08 00:00:00 EST 2001

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

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基因别名: ADMIO2; IMD48; mrtle; mur; SRK; STCD; STD; TZK; ZAP-70; ZAP70

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UniProt ID: (Human) P43403, (Mouse) P43404

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Entrez Gene ID: (Human) 7535, (Mouse) 22637

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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

参与通路

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