Invitrogen™ VISTA Monoclonal Antibody (MH5A), PerCP-eFluor™ 710, eBioscience™

Catalog No.	Quantity
46-591-982	100 μg

Catalog No. 46-591-982 Supplier Invitrogen™ Supplier No. 46591982

Description

Armenian Hamster Monoclonal Antibody

The monoclonal antibody MH5A reacts with a homolog of PD-1 (PD-1H) also known as VISTA. PD-1H is a 309 aa type I transmembrane protein that belongs to the CD28 superfamily. This cell surface receptor is expressed on nearly all hematopoietic cells except B cells. VISTA negatively regulates T cell responses by suppressing early TCR activation, inhibiting cell proliferation, and suppressing TH1 cytokine production. The MH5A antibody has been reported to inhibit T cell infiltration in graft-versus-host disease mouse models. Applications Reported: This MH5A antibody has been reported for use in flow cytometric analysis. Applications Tested: This MH5A antibody has been tested by flow cytometric analysis of mouse splenocytes. This can be used at less than or equal to 1 μg per test. A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 μL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. PerCP-eFluor 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome. Fixation: Samples can be stored in IC Fixation Buffer (Product #...

VISTA (V-domain Ig containing suppressor of T-cell activation), also known as B7-H5 or PD-1H, is a product of the VSIR gene. Although VISTA bears sequence homology to PD-L1 it has a different expression pattern. In both mouse and humans, VISTA is expressed predominantly on hematopoietic cells with the highest densities on myeloid cells, and lower levels on T cells. It is usually absent on B cells. VISTA expressed by antigen presenting cells has been shown to directly suppress proliferation and cytokine production of CD4 and CD8 T cells, making it a target for cancer immunotherapy. The reported ligands of VISTA include CD28H, VSIG-3 and VSIG-8.

Specifications

• Antigen: VISTA
• Applications: Flow Cytometry
• Classification: Monoclonal
• Clone: MH5A
• Concentration: 0.2 mg/mL
• Conjugate: PerCP-eFluor 710
• Formulation: PBS with 0.09% sodium azide; pH 7.2
• Gene: VSIR
• Gene Accession No.: Q9D659
• Gene Alias: 4632428N05Rik; B7H5; B7-H5; C10orf54; C1H10orf54; C28H10orf54; C4H10orf54; chromosome 10 open reading frame 54; DD1alpha; Death Domain1alpha; Dies1; differentiation of ESCs 1; GI24; PD-1H; PDCD1 homolog; PD-H1; platelet receptor Gi24; PP2135; RIKEN cDNA 4632428N05 gene; SISP1; sisp-1; Stress-induced secreted protein-1; UNQ730/PRO1412; V-domain Ig suppressor of T cell activation; VISTA; V-set domain-containing immunoregulatory receptor; V-set immunoregulatory receptor; VSIR; V-type immunoglobulin domain-containing suppressor of T-cell activation
• Gene Symbols: VSIR
• Host Species: Armenian Hamster
• Purification Method: Affinity chromatography
• Quantity: 100 μg
• Regulatory Status: RUO
• Primary or Secondary: Primary
• Gene ID (Entrez): 74048
• Target Species: Mouse
• Content And Storage: 4°C, store in dark, DO NOT FREEZE!
• Product Type: Antibody
• Form: Liquid
• Isotype: IgG

Frequently Asked Questions (FAQs)

I am unable to analyze my cells stained with eFluor Organic Dyes today. What options do I have?

Our options will depend on the samples you are analyzing.

If cell viability is not critical, you can store your stained samples at 4 degrees C or on ice overnight in the dark and analyze the following day.

For samples stained with eFluor organic fluorochromes, we recommend that cells be suspended in 100 uL of Flow Cytometry Staining Buffer (Cat. No. 00-4222) and 100 uL of eBioscience IC Fixation Buffer (Cat. No. 00-8222); samples can be incubated for up to 3 days at 4 degrees C in the dark. Alternatively, the 1-step Fix/Lyse Solution (Cat. No. 00-5333) can be used. This is a great option when working with whole blood but also works for other cell types.

Can the eFluor Organic fluorochromes be used for intracellular staining?

Yes, the eFluor Organic fluorochromes can be used for intracellular staining. The eFluor organic fluorochromes maintain bright signal and require minimal changes in compensation when fixed with eBioscience IC Fixation Buffer (Cat. No. 00-8222-49) and Permeabilization Buffer (Cat. No. 00-8333-56) or 1-step Fix/Lyse Solution (Cat. No. 00-5333-54, 00-5333-57) (as compared to live cells).

What buffers are compatible with the eFluor Organic fluorochromes and eVolve QDots?

The eFluor Organic fluorochromes and eVolve QDots can be used with flow staining buffers containing PBS and protein.

What is the difference between eFluor Organic Dyes and eVolve Qdots?

The eFluor Organic Dyes (eFluor 450, APC-eFluor 780, PerCP-eFluor 710, eFluor 710) are conventional fluorochromes. In contrast, the eVolve line of products are Quantum dots.

Are the eFluor Organic Dyes photo-labile?

As with other fluorochromes, we recommend minimal exposure to light to maintain optimal signal.