Revision 1
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, FC-FP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

76

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q13094

Entrez-Gene Id:

3937

Product Information

Product Usage Information

This product is the carrier free version of product #92711. All data were generated using the same antibody clone in the standard formulation which contains BSA and glycerol.

This formulation is ideal for use with technologies requiring specialized or custom antibody labeling, including fluorophores, metals, lanthanides, and oligonucleotides. It is not recommended for ChIP, ChIP-seq, CUT&RUN or CUT&Tag assays. If you require a carrier free formulation for chromatin profiling, please contact us. Optimal dilutions/concentrations should be determined by the end user.

Formulation

Supplied in 1X PBS, BSA and Azide Free.

For standard formulation of this product see product #92711

Storage

Store at -20°C. This product will freeze at -20°C so it is recommended to aliquot into single-use vials to avoid multiple freeze/thaw cycles. A slight precipitate may be present and can be dissolved by gently vortexing. This will not interfere with antibody performance.

Specificity / Sensitivity

Phopho-SLP-76 (Ser376) (D7S1K) Rabbit mAb recognizes endogenous levels of SLP-76 protein only when phosphorylated at Ser376. This product reacts with mouse by western blot only. Clone E3G9U is more sensitive by flow cytometry than clone D7S1K.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser376 of human SLP-76 protein.

Background

SH2 domain-containing leukocyte protein of 76 kDa (SLP-76) is a hematopoietic adaptor protein that is important in multiple biochemical signaling pathways and necessary for T cell development and activation (1). ZAP-70 phosphorylates SLP-76 and LAT as a result of TCR ligation. SLP-76 has amino-terminal tyrosine residues followed by a proline-rich domain and a carboxy-terminal SH2 domain. Phosphorylation of Tyr113 and Tyr128 result in recruitment of the GEF Vav and the adaptor protein Nck (2). TCR ligation also leads to phosphorylation of Tyr145, which mediates an association between SLP-76 and Itk, which is accomplished in part via the proline-rich domain of SLP-76 and the SH3 domain of Itk (3). Furthermore, the proline-rich domain of SLP-76 binds to the SH3 domains of Grb2-like adaptor Gads (3,4). In resting cells, SLP-76 is predominantly in the cytosol. Upon TCR ligation, SLP-76 translocates to the plasma membrane and promotes the assembly of a multi-protein signaling complex that includes Vav, Nck, Itk, and PLCγ1 (1). The expression of SLP-76 is tightly regulated; the protein is detected at very early stages of thymocyte development, increases as thymocyte maturation progresses, and is reduced as cells mature to CD4+ CD8+ double-positive thymocytes (5).

Following TCR ligation, SLP-76 is phosphorylated at Ser376 by the hematopoietic progenitor kinase 1 (HPK1) (6,7). This phosphorylation induces interaction with 14-3-3ε, which leads to the disassembly of TCR signaling complexes and downregulation of TCR signaling (6-8).

  1. Clements, J.L. (2003) Immunol Rev 191, 211-9.
  2. Bubeck Wardenburg, J. et al. (1998) Immunity 9, 607-16.
  3. Bunnell, S.C. et al. (2000) J Biol Chem 275, 2219-30.
  4. Liu, S.K. et al. (1999) Curr Biol 9, 67-75.
  5. Clements, J.L. et al. (1998) J Immunol 161, 3880-9.
  6. Shui, J.W. et al. (2007) Nat Immunol 8, 84-91.
  7. Di Bartolo, V. et al. (2007) J Exp Med 204, 681-91.
  8. Lasserre, R. et al. (2011) J Cell Biol 195, 839-53.

Species Reactivity

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

Applications Key

WB: Western Blotting 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 1
#72511

Phospho-SLP-76 (Ser376) (D7S1K) Rabbit mAb (BSA and Azide Free)

Western Blotting Image 1: Phospho-SLP-76 (Ser376) (D7S1K) Rabbit mAb (BSA and Azide Free) Expand Image
使用 Phospho-SLP-76 (Ser376) (D7S1K) Rabbit mAb(上图)、Rabbit mAb (upper), SLP-76 (D1R1A) Rabbit mAb #70896(中图)或 β-Actin (D6A8) Rabbit mAb #8457 (下图),对未经处理 (-) 或已经 (+) H2O2 (11 mM,1 分钟)或交联抗 CD3 和抗 CD28(各 10 μg/mL,15 分钟)处理的血清饥饿 Jurkat 细胞的提取物进行蛋白质印迹法分析。数据的生成使用的是该产品的常规组分产品。
Western Blotting Image 2: Phospho-SLP-76 (Ser376) (D7S1K) Rabbit mAb (BSA and Azide Free) Expand Image
使用 Phospho-SLP-76 (Ser376) (D7S1K) Rabbit mAb(上图)、SLP-76 (D1R1A) Rabbit mAb #70896(中图)或 β-Actin (D6A8) Rabbit mAb #8457 (下图),对未经处理 (-) 或已经牛小肠磷酸酶 (CIP) 和 λ 磷酸酶 (+) 处理的 Jurkat 细胞的提取物进行蛋白质印迹法分析。数据的生成使用的是该产品的常规组分产品。
Flow Cytometry Image 1: Phospho-SLP-76 (Ser376) (D7S1K) Rabbit mAb (BSA and Azide Free) Expand Image
使用 Phospho-SLP-76 (Ser376) (D7S1K) Rabbit mAb(顶排)或浓度匹配的Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (底排)与共染 CD3 抗体对未经处理(左列)或已经交联抗 CD3 和抗 CD28(各 10 μg/ml,15 分钟;右列)处理的人外周血单核细胞进行流式细胞分析。Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 用作二抗。数据的生成使用的是该产品的常规组分产品。