Thermo Fisher Scientific VISTA Monoclonal Antibody (MIH64), NovaFluor Yellow 730, eBioscience

Thermo Fisher Scientific VISTA Monoclonal Antibody (MIH64), NovaFluor Yellow 730, eBioscience

Applications

• Flow Cytometry (Flow)

Tested Dilution: 0.4 µg/test

Product Specifications

Additional Conjugate Formats

• APC
• Brilliant UV 615
• Brilliant UV 737
• NovaFluor Blue 660-120S
• NovaFluor Yellow 610
• PE
• PerCP-eFluor 710
• Super Bright 436
• Super Bright 600
• Request custom conjugation available

Product Specific Information

This MIH64 monoclonal antibody recognizes mouse VISTA (B7-H5 or PD-H1).
It is suitable for flow cytometry on both native and paraformaldehyde-fixed cells.

Each product contains:

• 1 vial of NovaFluor conjugate
• 1 vial of CellBlox Plus Blocking Buffer

Applications Reported: Flow cytometric analysis
Applications Tested: Flow cytometric analysis of mouse splenocytes at ≤ 0.4 µg/test (100 µL final volume).
Cell number should be empirically determined (10⁵–10⁸ cells/test).
Careful titration is recommended for optimal performance.

Important Notes:

• NovaFluor dyes are not compatible with DNA intercalating viability dyes (e.g., propidium iodide, 7-AAD, DAPI).
• Use Invitrogen LIVE/DEAD Fixable Dead Cell stains for viability detection.
• CellBlox Plus Blocking Buffer (Cat. No. C001T06F01) is required for optimal staining with NovaFluor conjugates.
  • Add 5 µL per sample, regardless of the number of NovaFluors used.
  • For single-color controls, use 5 µL per 100 µL of cell sample (10³–10⁸ cells).

NovaFluor conjugates utilize Phiton™ technology, based on nucleic acid dye structures engineered to minimize spillover and spread.
Excitation: 552 nm; Emission: 718 nm; Laser: 561 nm (Yellow)

Note: Internal testing shows NovaFluor Yellow 730 may stain B cells non-specifically in SJL mice.
No non-specific staining observed in BALB/c or C57BL/6 strains. Other strains not tested.

Target Information

VISTA (V-domain Ig containing suppressor of T-cell activation), also known as B7-H5 or PD-1H, is encoded by the VSIR gene.
It shares homology with PD-L1 but exhibits distinct expression patterns.
In both mouse and humans, VISTA is predominantly expressed on hematopoietic cells—highest on myeloid cells, lower on T cells, and typically absent on B cells.
VISTA expressed by antigen-presenting cells suppresses CD4 and CD8 T-cell proliferation and cytokine production, making it a potential target for cancer immunotherapy.
Reported ligands include CD28H, VSIG-3, and VSIG-8.

For Research Use Only.
Not for use in diagnostic procedures or resale without authorization.

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