Revision 1

#48287Store at -20C

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

REACTIVITY:

H M

SENSITIVITY:

Endogenous

MW (kDa):

40

SOURCE:

Rabbit

UniProt ID:

#O75791

Entrez-Gene Id:

9402

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000

Storage

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

Specificity / Sensitivity

GADS Antibody recognizes endogenous levels of total GADS protein.

Species Reactivity:

Human, Mouse

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly172 of human GADS protein. Antibodies are purified by peptide affinity chromatography.

Background

GRB2-related adaptor downstream of Shc (GADS) belongs to the GRB2 family of adaptor proteins. It is a hematopoietic cell-specific signaling adaptor protein that harbors amino- and carboxy-terminal SH3 domains, a central SH2 domain, and a unique linker region that is rich in proline and glutamine residues (1). The presence of SH2 and SH3 domains within GADS strongly suggest that it functions in signal transduction cascades by facilitating protein-protein interactions. In the context of T cells, research studies have demonstrated that GADS interacts with LAT and SLP-76 signaling complexes to facilitate NFAT activation downstream of TCR engagement (2-4). Given its role as a fundamental mediator of TCR signaling, GADS is subject to multiple modes of negative regulation to limit TCR signal strength. For example, research studies have demonstrated that HPK1 directly phosphorylates GADS at Thr262 within its linker region, a modification that promotes 14-3-3 binding and dissociation of signaling complexes nucleated by LAT, SLP-76, and GADS (5). The linker region of GADS is also subject to caspase-mediated cleavage, which separates its SH2 and SH3 domains and thus impairs the ability of GADS to bridge LAT and SLP-76 signaling complexes for transduction of faithful TCR signals (6,7).

  1. Hámori, J. et al. (1978) Neuroscience 3, 403-12.
  2. Liu, S.K. et al. (1999) Curr Biol 9, 67-75.
  3. Law, C.L. et al. (1999) J Exp Med 189, 1243-53.
  4. Asada, H. et al. (1999) J Exp Med 189, 1383-90.
  5. Lasserre, R. et al. (2011) J Cell Biol 195, 839-53.
  6. Berry, D.M. et al. (2001) Oncogene 20, 1203-11.
  7. Yankee, T.M. et al. (2001) Proc Natl Acad Sci USA 98, 6789-93.

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 nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting

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
#48287

GADS Antibody

Western Blotting Image 1: GADS Antibody Expand Image
使用 GADS Antibody(上图)或 β-Actin (D6A8) Rabbit mAb #8457(下图)对各种人细胞系的提取物进行蛋白质印迹分析。GADS 蛋白在 Ramos 细胞中阴性表达与已发表的观察结果一致。
Western Blotting Image 2: GADS Antibody Expand Image
使用 GADS Antibody(上图)、Myc-Tag (71D10) Rabbit mAb #2278(中图)和 β-Actin (D6A8) Rabbit mAb #8457(下图)对空载转染 (-) 或经表达 Myc/DDK 标记的全长人 GADS (hGADS-Myc/DDK; +) 的构建体转染的 293T 细胞的提取物进行蛋白质印迹分析。