WB result of VISTA Recombinant Rabbit mAb
Primary antibody: VISTA Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: MCF7 whole cell lysate 20 µg
Lane 2: HeLa whole cell lysate 20 µg
Lane 3: DU 145 whole cell lysate 20 µg
Lane 4: Jurkat whole cell lysate 20 µg
Negative control: MCF7 whole cell lysate
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 34 kDa
Observed MW: 40~55 kDa
VISTA Recombinant Rabbit mAb (S-1469-72)
VISTA Recombinant Rabbit mAb (S-1469-72)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | VISTA |
| Synonyms | V-type immunoglobulin domain-containing suppressor of T-cell activation; Platelet receptor Gi24; Stress-induced secreted protein-1 (Sisp-1); V-set domain-containing immunoregulatory receptor; V-set immunoregulatory receptor; VSIR; C10orf54; SISP1 |
| Immunogen | Recombinant Protein |
| Location | Cell membrane |
| Accession | Q9H7M9 |
| Clone Number | S-1469-72 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, IP, FCM |
| Reactivity | Hu |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:1000 | |
| IP | 1:50 | |
| IHC-P | 1:250 | |
| FCM | 1:50 |
Background
VISTA is an important immune modulatory receptor that is gaining attention in cancer immunotherapy. VISTA exerts its suppressive effects on T cell activity by binding to their respective ligands on T cells, potentially influencing the immune evasion mechanisms of tumors. Studies have shown that VISTA is expressed in a variety of cancers, including melanoma, pancreatic cancer, and prostate cancer, and its expression levels are closely related to the immunosuppressive state of tumors.
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Western Blot
FC
Flow cytometric analysis of human PBMC (human peripheral blood mononuclear cell) labelling VISTA antibody at 1/50 dilution (1 μg) / (Red) compared with a Rabbit monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti - Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody. Then cells were stained with CD14 - Alexa Fluor® 488 separately. Gated on total viable cells.
IP
VISTA Rabbit mAb at 1/50 dilution (1 µg) immunoprecipitating VISTA in 0.4 mg DU 145 whole cell lysate.
Western blot was performed on the immunoprecipitate using VISTA Rabbit mAb at 1/1000 dilution.
Secondary antibody (HRP) for IP was used at 1/1000 dilution.
Lane 1: DU 145 whole cell lysate 20 µg (Input)
Lane 2: VISTA Rabbit mAb IP in DU 145 whole cell lysate
Lane 3: Rabbit monoclonal IgG IP in DU 145 whole cell lysate
Predicted MW: 34 kDa
Observed MW: 40~55 kDa
This blot was developed with high sensitivity substrate
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human placenta. Anti-VISTA antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human tonsil. Anti-VISTA antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human liver. Anti-VISTA antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Negative control: IHC shows negative staining in paraffin-embedded human skeletal muscle. Anti-VISTA antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human endometrial cancer. Anti-VISTA antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human cervical squamous cell carcinoma. Anti-VISTA antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human colon cancer. Anti-VISTA antibody was used at 1/250 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
