Mouse VISTA/B7-H5/PD-1H Antibody

  
  • Species Reactivity
    Mouse
  • Specificity
    Detects mouse VISTA/B7-H5/PD-1H in direct ELISAs and Western blots. In direct ELISAs, approximately 25% cross-reactivity with recombinant human VISTA/B7-H5/PD-1H is observed.
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant mouse VISTA/B7-H5/PD-1H
    Phe33-Ala191
    Accession # Q9D659
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Flow Cytometry
    0.25 µg/106 cells
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    5-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of VISTA/B7-H5/PD-1H by Western Blot. Western blot shows lysates of mouse platelets. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Mouse VISTA/B7-H5/PD-1H Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7005) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for VISTA/B7-H5/PD-1H at approximately 30 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Detection of VISTA/B7-H5/PD-1H in HEK293 Human Cell Line Transfected with Mouse VISTA and eGFP by Flow Cytometry. HEK293 human embryonic kidney cell line transfected with either
(A) mouse VISTA or (B) irrelevant transfectants and eGFP was stained with Sheep Anti-Mouse VISTA/B7-H5/PD-1H Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7005) followed by Allophycocyanin-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # F0127). Quadrant markers were set based on control antibody staining (Catalog # 5-001-A). View our protocol for Staining Membrane-associated Proteins.
Immunocytochemistry
VISTA/B7-H5/PD-1H in D3 Mouse Cell Line. VISTA/B7-H5/PD-1H was detected in immersion fixed D3 mouse embryonic stem cell line using Sheep Anti‑Mouse VISTA/B7-H5/PD-1H Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7005) at
10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
  • Reconstitution
    Sterile PBS to a final concentration of 0.2 mg/mL.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: VISTA/B7-H5/PD-1H

Platelet Receptor Gi24 (also known as VISTA, B7-H5, SISP1, C10orf54 and, Dies1 [Differentiation of ESC-1]) is a 55-65 kDa member of the Ig superfamily. It is a transmembrane molecule expressed in bone, on embryonic stem cells (ESCs), and on tumor cell surfaces. On ESCs, Gi24 appears to positively interact with BMP4, potentiating BMP signaling and the transition from an undifferentiated to a differentiated state. On tumor cells, Gi24 both promotes MT1-MMP expression and activity, and serves as a substrate for MT1-MMP. This increases the potential for cell motility. Mature mouse Gi24 is a 277 amino acid (aa) type I transmembrane glycoprotein (aa 33-309). It contains a 149 aa extracellular region (aa 33-191) with one V-type Ig-like domain (aa 33-161) and a 97 aa cytoplasmic domain. Based on human Gi24, mouse Gi24 will likely undergo proteolytic cleavage by MT1-MMP, generating a soluble 30 kDa extracellular fragment, plus a 25-30 kDa membrane-bound fragment. There are two potential isoform variants. One contains a deletion of aa 127-187, while another shows an alternative start site at Met82. Over aa 33-191, mouse Gi24/Dies1 shares 78% and 70% aa identity with rat and human Gi24, respectively.

  • Entrez Gene IDs:
    64115 (Human); 74048 (Mouse); 690899 (Rat); 102116468 (Cynomolgus Monkey)
  • Alternate Names:
    4632428N05Rik; B7H5; B7-H5; C10orf54; chromosome 10 open reading frame 54; Dies1; Gi24; PD1H; PD-1H; platelet receptor Gi24; PP2135; SISP1; stress induced secreted protein 1; VISTA
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