Revision 18

#99567Store 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, IHC-P, IF-IC

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

18, 30-40

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q495A1

Entrez-Gene Id:

201633

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:2000
Immunohistochemistry (Paraffin) 1:200 - 1:800
Immunofluorescence (Immunocytochemistry) 1:800 - 1:3200

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 #17046.

Specificity / Sensitivity

TIGIT (E5Y1W) XP® Rabbit mAb recognizes endogenous levels of total TIGIT protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the carboxy terminus of human TIGIT protein. This antibody cross-reacts with an unidentified protein of 42 kDa in some cell extracts.

Background

T-cell immunoreceptor with Ig and ITIM domains (TIGIT), also known as VSIG9, VSTM3, and WUCAM, is a member of the poliovirus receptor family of immunoglobulin proteins (1-3). TIGIT is expressed at low levels on subsets of T cells and NK cells, and is upregulated at the protein level following activation of these cells (1-4). TIGIT marks exhausted T cells in the tumor microenvironment (5) and during human immunodeficiency virus (HIV) infection (6). Research has shown TIGIT interacts with several receptors expressed on antigen presenting cells, such as dendritic cells and macrophages, as well as tumor cells and cells of the microenvironment. TIGIT binds with high affinity to PVR/CD155, and with low affinity to Nectin-2/CD112 and Nectin-3/CD113 (2,4,7). Upon binding to its ligands, TIGIT suppresses T cell activation, and inhibits T and NK cell cytotoxicity. This inhibition can be blocked using monoclonal antibodies directed at the extracellular domain of TIGIT, resulting in rejuvenated antigen-specific CD8+ T cell responses in tumors and during HIV infection (5,6,8). Three potential isoforms of TIGIT have been computationally mapped (9).

  1. Yu, X. et al. (2009) Nat Immunol 10, 48-57.
  2. Levin, S.D. et al. (2011) Eur J Immunol 41, 902-15.
  3. Boles, K.S. et al. (2009) Eur J Immunol 39, 695-703.
  4. Stanietsky, N. et al. (2009) Proc Natl Acad Sci U S A 106, 17858-63.
  5. Chauvin, J.M. et al. (2015) J Clin Invest 125, 2046-58.
  6. Chew, G.M. et al. (2016) PLoS Pathog 12, e1005349.
  7. Stengel, K.F. et al. (2012) Proc Natl Acad Sci U S A 109, 5399-404.
  8. Johnston, R.J. et al. (2014) Cancer Cell 26, 923-37.
  9. Bechtel, S. et al. (2007) BMC Genomics 8, 399.

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 IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry)

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.
SignalStain is a registered trademark of Cell Signaling Technology, Inc.
XP is a registered 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 18
#99567

TIGIT (E5Y1W) XP® Rabbit mAb

Western Blotting Image 1: TIGIT (E5Y1W) XP® Rabbit mAb Expand Image
使用 TIGIT (E5Y1W) XP® Rabbit mAb(上图)和 β-Actin (D6A8) Rabbit mAb #8457 (下图)对人 CD8+ T 细胞以及不同细胞系的提取物进行蛋白质印迹分析。
Immunohistochemistry Image 1: TIGIT (E5Y1W) XP® Rabbit mAb Expand Image
使用 TIGIT (E5Y1W) XP® Rabbit mAb 对石蜡包埋的 293T 细胞沉淀物、对照物(左上图)或 TIGIT 转染物(右上图),MJ [G11] 细胞沉淀物(左下图,阳性)和纯化的 CD8+ 人外周血单核细胞沉淀物(右下图,阳性)进行免疫组织化学分析。使用人血纯化 CD8+ T 细胞,并使用存在human interleukin-2 (hIL-2) #8907 (7 ng/ml) 的 CD3 和 CD28 抗体涂层球刺激 7 天。
Immunohistochemistry Image 2: TIGIT (E5Y1W) XP® Rabbit mAb Expand Image
使用 TIGIT (E5Y1W) XP® Rabbit mAb 对石蜡包埋的人尿路上皮癌进行免疫组织化学分析。
Immunohistochemistry Image 3: TIGIT (E5Y1W) XP® Rabbit mAb Expand Image
使用 TIGIT (E5Y1W) XP® Rabbit mAb 对石蜡包埋的人结肠腺癌进行免疫组织化学分析。
Immunohistochemistry Image 4: TIGIT (E5Y1W) XP® Rabbit mAb Expand Image
使用 TIGIT (E5Y1W) XP® Rabbit mAb 对石蜡包埋的人食管癌进行免疫组织化学分析。
Immunohistochemistry Image 5: TIGIT (E5Y1W) XP® Rabbit mAb Expand Image
使用 TIGIT (E5Y1W) XP® Rabbit mAb 对石蜡包埋的人鳞状细胞癌进行免疫组织化学分析。
Immunohistochemistry Image 6: TIGIT (E5Y1W) XP® Rabbit mAb Expand Image
使用 TIGIT (E5Y1W) XP® Rabbit mAb 对石蜡包埋的人非小细胞肺癌进行免疫组织化学分析。
Immunohistochemistry Image 7: TIGIT (E5Y1W) XP® Rabbit mAb Expand Image
以浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(右图)作为对照,使用 TIGIT (E5Y1W) XP® Rabbit mAb(左图)对石蜡包埋的人 B 细胞非霍奇金淋巴瘤进行免疫组织化学分析。
Immunohistochemistry Image 8: TIGIT (E5Y1W) XP® Rabbit mAb Expand Image
使用 TIGIT (E5Y1W) XP® Rabbit mAb 对石蜡包埋的人前列腺癌进行免疫组织化学分析。
Immunohistochemistry Image 9: TIGIT (E5Y1W) XP® Rabbit mAb Expand Image
使用 TIGIT (E5Y1W) XP® Rabbit mAb 对石蜡包埋的人扁桃体进行免疫组织化学分析。
Immunofluorescence Image 1: TIGIT (E5Y1W) XP® Rabbit mAb Expand Image
使用 TIGIT (E5Y1W) XP® Rabbit mAb(绿色)和 CD8α (RPA-T8) Mouse mAb (FITC Conjugate) #55397(伪彩红色)对人 CD8+ T 细胞进行共聚焦免疫荧光分析。样本用 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)封片。使用人血纯化 CD8+ T 细胞,并使用存在human interleukin-2 (hIL-2) #8907 (20 ng/ml) 的 CD3 和 CD28 抗体涂层球刺激 7 天。
Immunofluorescence Image 2: TIGIT (E5Y1W) XP® Rabbit mAb Expand Image
使用 TIGIT (E5Y1W) XP® Rabbit mAb(绿色)对 MJ [G11] 细胞(左图,阳性)和 IGROV-1 细胞(右图,阴性)进行共聚焦免疫荧光分析。样本用 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)封片。