Revision 2
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-Bond, IHC-P, FC-FP

REACTIVITY:

H Mk

SENSITIVITY:

Endogenous

MW (kDa):

45-65

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q9H7M9

Entrez-Gene Id:

64115

Product Information

Product Usage Information

This product is the carrier-free version of product #64953. 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 #64953.

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

VISTA (D1L2G) XP® Rabbit mAb (BSA and Azide Free) recognizes endogenous levels of total VISTA protein. Non-specific staining was observed in kidney and prostate epithelium by immunohistochemistry.

Species Reactivity:

Human, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human 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).

Applications Key

WB: Western Blotting IHC-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) 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.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 2
#82119

VISTA (D1L2G) XP® Rabbit mAb (BSA and Azide Free)

Western Blotting Image 1: VISTA (D1L2G™) XP® Rabbit mAb (BSA and Azide Free) Expand Image
Western blot analysis of extracts from OVKATE, DU 145, and MCF7 cells using VISTA ((D1L2G) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). Data were generated using the standard formulation of this product.
Immunohistochemistry Image 1: VISTA (D1L2G™) XP® Rabbit mAb (BSA and Azide Free) Expand Image
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using VISTA (D1L2G) XP® Rabbit mAb performed on the Leica® BOND Rx. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 1: VISTA (D1L2G™) XP® Rabbit mAb (BSA and Azide Free) Expand Image
Immunohistochemical analysis of paraffin-embedded human tonsil using VISTA (D1L2G) XP® Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 2: VISTA (D1L2G™) XP® Rabbit mAb (BSA and Azide Free) Expand Image
Immunohistochemical analysis of paraffin-embedded human non-Hodgkin B-Cell lymphoma using VISTA (D1L2G) XP® Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 3: VISTA (D1L2G™) XP® Rabbit mAb (BSA and Azide Free) Expand Image
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using VISTA (D1L2G) XP® Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 4: VISTA (D1L2G™) XP® Rabbit mAb (BSA and Azide Free) Expand Image
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using VISTA (D1L2G) XP® Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 5: VISTA (D1L2G™) XP® Rabbit mAb (BSA and Azide Free) Expand Image
Immunohistochemical analysis of paraffin-embedded rhesus monkey spleen using VISTA (D1L2GTM) XP® Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 6: VISTA (D1L2G™) XP® Rabbit mAb (BSA and Azide Free) Expand Image
Immunohistochemical analysis of paraffin-embedded OVKATE (left) and MCF7 (right) cell pellets using VISTA (D1L2G) XP® Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 7: VISTA (D1L2G™) XP® Rabbit mAb (BSA and Azide Free) Expand Image
Multiplex immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using VISTA (D1L2G) XP® Rabbit mAb (orange), PD-1 (D4W2J) XP® Rabbit mAb #86163 (green), B7-H3 (D9M2L) XP® Rabbit mAb #14058 (red), B7-H4 (D1M8I) XP® Rabbit mAb (cyan), LAG3 (D2G4O) XP® Rabbit mAb #15372 (magenta), and TIM-3 (D5D5R) XP® Rabbit mAb #45208 (yellow). Data were generated using the standard formulation of this product.
Flow Cytometry Image 1: VISTA (D1L2G™) XP® Rabbit mAb (BSA and Azide Free) Expand Image
Flow cytometric analysis of fixed and permeabilized human whole blood cells stained with VISTA (D1L2G) XP® Rabbit mAb or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 and co-stained with CD19-PE and CD14-APC. VISTA is present on CD14+ monocytes (right; upper right quadrant) and shows a significant increase in intensity when compared to its isotype control (middle; upper left quadrant). CD19+ B cells are negative for VISTA (left; upper left quadrant). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. Data were generated using the standard formulation of this product.