Revision 6
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IF-IC, FC-FP

REACTIVITY:

H M

SENSITIVITY:

Endogenous

MW (kDa):

70, 72

Source/Isotype:

Rabbit IgG

UniProt ID:

#P43403, #P43405

Entrez-Gene Id:

7535, 6850

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunofluorescence (Immunocytochemistry) 1:200 - 1:800
Flow Cytometry (Fixed/Permeabilized) 1:400 - 1:1600

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #31754.

Specificity / Sensitivity

Phospho-Zap-70 (Tyr319)/Syk (Tyr352) (65E4) Rabbit mAb detects endogenous levels of Zap-70 only when phosphorylated at Tyr319. It cross-reacts with endogenous levels of Syk when phosphorylated at Tyr352.

Species Reactivity:

Human, Mouse

Species predicted to react based on 100% sequence homology

Rat, Hamster, Chicken, Bovine, Dog, Pig, Horse

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr319 of human Zap-70.

Background

The Syk family protein tyrosine kinase Zap-70 is expressed in T and NK cells and plays a critical role in mediating T cell activation in response to T cell receptor (TCR) engagement (1). Following TCR engagement, Zap-70 is rapidly phosphorylated on several tyrosine residues through autophosphorylation and transphosphorylation by the Src family tyrosine kinase Lck (2-6). Tyrosine phosphorylation correlates with increased Zap-70 kinase activity and downstream signaling events. Expression of Zap-70 is correlated with disease progression and survival in patients with chronic lymphocytic leukemia (7,8).

  1. Chu, D.H. et al. (1998) Immunol Rev 165, 167-80.
  2. Iwashima, M. et al. (1994) Science 263, 1136-9.
  3. Neumeister, E.N. et al. (1995) Mol Cell Biol 15, 3171-8.
  4. Chan, A.C. et al. (1995) EMBO J 14, 2499-508.
  5. Williams, B.L. et al. (1999) EMBO J 18, 1832-44.
  6. Di Bartolo, V. et al. (1999) J Biol Chem 274, 6285-94.
  7. Wiestner, A. et al. (2003) Blood 101, 4944-51.
  8. Crespo, M. et al. (2003) N Engl J Med 348, 1764-75.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 6
#2717

Phospho-Zap-70 (Tyr319)/Syk (Tyr352) (65E4) Rabbit mAb

Western Blotting Image 1: Phospho-Zap-70 (Tyr319)/Syk (Tyr352) (65E4) Rabbit mAb Expand Image
使用 Phospho-Zap-70 (Tyr319)/Syk (Tyr352) (65E4) Rabbit mAb 对已经过氧化氢(2mM,2 分钟)或 λ 磷酸酶处理的 Jurkat 细胞的提取物以及已经抗人 IgM(12 mcg/ml,2 分钟)处理的 Ramos 细胞的提取物进行蛋白质印迹分析。
Immunofluorescence Image 1: Phospho-Zap-70 (Tyr319)/Syk (Tyr352) (65E4) Rabbit mAb Expand Image
使用 Phospho-Zap-70 (Tyr319)/Syk (Tyr352) (65E4) Rabbit mAb(绿色)对未经处理(右图)或已经人 IgM(左图)处理的 Ramos 细胞进行共聚焦免疫荧光分析。红色 = Propidium Iodide (PI)/RNase Staining Solution #4087。
Flow Cytometry Image 1: Phospho-Zap-70 (Tyr319)/Syk (Tyr352) (65E4) Rabbit mAb Expand Image
使用 Phospho-Zap-70 (Tyr319)/Syk (Tyr352) (65E4) Rabbit mAb(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)对未经处理(蓝色)或已经过抗 IgM(12 ug/ml,2 分钟;绿色)处理的 Ramos 细胞进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 作为二抗。