Revision 4

#54979

Store at -20C

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, 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 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.

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).

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

W: 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

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

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 4

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines and tissues using VISTA (D5L5T) XP® Rabbit mAb (upper) and β-actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 1: VISTA (D5L5T) XP<sup>®</sup> Rabbit mAb
Immunoprecipitation of VISTA protein from EL4 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is VISTA (D5L5T) XP® Rabbit mAb. Western blot analysis was performed using VISTA (D5L5T) XP® Rabbit mAb.
Immunoprecipitation Image 1: VISTA (D5L5T) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded mouse CT26 syngeneic tumor using VISTA (D5L5T) XP® Rabbit mAb performed on the Leica® Bond™ Rx.
Immunohistochemistry Image 1: VISTA (D5L5T) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 4

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded mouse colon using VISTA (D5L5T) XP® Rabbit mAb.
Immunohistochemistry Image 1: VISTA (D5L5T) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded mouse lung using VISTA (D5L5T) XP® Rabbit mAb.
Immunohistochemistry Image 2: VISTA (D5L5T) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded EL4 cell pellet (left, positive) or 3T3 cell pellet (right, negative) using VISTA (D5L5T) XP® Rabbit mAb.
Immunohistochemistry Image 3: VISTA (D5L5T) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 4

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded mouse spleen using VISTA (D5L5T) XP® Rabbit mAb.
Immunohistochemistry Image 4: VISTA (D5L5T) XP<sup>®</sup> Rabbit mAb
Confocal immunofluorescent analysis of mouse colon using VISTA (D5L5T) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).
Immunofluorescence Image 1: VISTA (D5L5T) XP<sup>®</sup> Rabbit mAb
Confocal immunofluorescent analysis of Raw 264.7 cells (left, positive) and HeLa cells (right, negative) using VISTA (D5L5T) XP® Rabbit mAb (green) and a β-Actin Mouse mAb (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunofluorescence Image 1: VISTA (D5L5T) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.