Revision 4

#54979Store 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, IP, IHC-Bond, IHC-P, IF-F, IF-IC

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

45-65

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q9D659

Entrez-Gene Id:

74048

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
IHC Leica Bond 1:300
Immunohistochemistry (Paraffin) 1:300
Immunofluorescence (Frozen) 1:200
Immunofluorescence (Immunocytochemistry) 1:200

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 #56548.

Specificity / Sensitivity

VISTA (D5L5T) XP® Rabbit mAb recognizes endogenous levels of total VISTA protein. This antibody is not recommended for immunohistochemical analysis of human tissues. Instead, VISTA (D1L2G) XP® Rabbit mAb #64953 is suggested.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Val291 of mouse VISTA protein.

Background

VISTA (V-Domain Ig Suppressor of T Cell Activation) is a negative checkpoint control protein that regulates T cell activation and immune responses. VISTA, which contains a single Ig-like V-type domain, a transmembrane domain, and an intracellular domain, has sequence similarity to both the B7 and CD28 family members. Although primarily expressed by myeloid cells, VISTA is also expressed by CD4+, CD8+, and FoxP3+ T-cells. Thus, VISTA is described as both a ligand and a receptor (1-3). Blocking VISTA induces T-cell activation and proliferation, and potentiates disease severity in the EAE model (1). Furthermore, genetic deletion of VISTA in mice leads to spontaneous T-cell activation and chronic inflammation (4,5). In mouse models of cancer, neutralization of VISTA enhances T-cell proliferation and effector function and increases tumor infiltration, suggesting VISTA blockade could be an effective strategy for tumor immunotherapy (6,7).

  1. Wang, L. et al. (2011) J Exp Med 208, 577-92.
  2. Flies, D.B. et al. (2011) J Immunol 187, 1537-41.
  3. Lines, J.L. et al. (2014) Cancer Res 74, 1924-32.
  4. Wang, L. et al. (2014) Proc Natl Acad Sci U S A 111, 14846-51.
  5. Liu, J. et al. (2015) Proc Natl Acad Sci U S A 112, 6682-7.
  6. Le Mercier, I. et al. (2014) Cancer Res 74, 1933-44.
  7. Lines, J.L. et al. (2014) Cancer Immunol Res 2, 510-7.

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 IP: Immunoprecipitation IHC-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) IF-F: Immunofluorescence (Frozen) 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.
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 4
#54979

VISTA (D5L5T) XP® Rabbit mAb

Western Blotting Image 1: VISTA (D5L5T) XP® Rabbit mAb Expand Image
使用 VISTA (D5L5T) XP® Rabbit mAb (上图)和 β-Actin (D6A8) Rabbit mAb #8457(下图),对不同细胞系和组织的提取物进行蛋白质印迹分析。
Immunoprecipitation Image 1: VISTA (D5L5T) XP® Rabbit mAb Expand Image
对 EL4 细胞提取物 VISTA 蛋白进行免疫沉淀。泳道 1 为 10% input,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 VISTA (D5L5T) XP® Rabbit mAb。使用 VISTA (D5L5T) XP® Rabbit mAb 进行蛋白质印迹分析。
Immunohistochemistry Image 1: VISTA (D5L5T) XP® Rabbit mAb Expand Image
在 Leica® Bond Rx 上使用 VISTA (D5L5T) XP® Rabbit mAb 对石蜡包埋的小鼠 CT26 同源肿瘤细胞进行免疫组织化学分析。
Immunohistochemistry Image 1: VISTA (D5L5T) XP® Rabbit mAb Expand Image
使用 VISTA (D5L5T) XP® Rabbit mAb 对石蜡包埋的小鼠结肠进行免疫组织化学分析。
Immunohistochemistry Image 2: VISTA (D5L5T) XP® Rabbit mAb Expand Image
使用 VISTA (D5L5T) XP® Rabbit mAb 对石蜡包埋的小鼠肺进行免疫组织化学分析。
Immunohistochemistry Image 3: VISTA (D5L5T) XP® Rabbit mAb Expand Image
使用 VISTA (D5L5T) XP® Rabbit mAb 对石蜡包埋的 EL4 细胞沉淀物(左图,阳性)或 3T3 细胞沉淀物(右图,阴性)进行免疫组织化学分析。
Immunohistochemistry Image 4: VISTA (D5L5T) XP® Rabbit mAb Expand Image
使用 VISTA (D5L5T) XP® Rabbit mAb 对石蜡包埋的小鼠脾细胞进行免疫组织化学分析。
Immunofluorescence Image 1: VISTA (D5L5T) XP® Rabbit mAb Expand Image
使用 VISTA (D5L5T) XP® Rabbit mAb(绿色)对小鼠结肠进行共聚焦免疫荧光分析。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。肌动蛋白微丝由 DyLight 554 Phalloidin #13054(红色)标记。
Immunofluorescence Image 1: VISTA (D5L5T) XP® Rabbit mAb Expand Image
使用 VISTA (D5L5T) XP® Rabbit mAb(绿色)和一种 β-Actin Mouse mAb(红色)对 Raw 264.7(左图,阳性)细胞和 Hela(右图,阴性)细胞进行共聚焦免疫荧光分析。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。