Thermo Fisher Scientific CD8a Monoclonal Antibody (53-6.7), NovaFluor Yellow 700, eBioscience

Applications

• Flow Cytometry (Flow): Tested at ≤ 0.5 µg/test

Product Specifications

Additional Available Formats

Unconjugated, Alexa Fluor 488, 532, 561, 660, 700, APC, APC-eFluor 780, Biotin, Brilliant UV/Violet series, eFluor series, FITC, PE, PE-Cyanine series, PerCP series, Super Bright series, NovaFluor Blue/Red/UV/Violet/Yellow series, and custom conjugation available.

Product Specific Information

The 53-6.7 monoclonal antibody reacts with the mouse CD8a molecule (32–34 kDa). CD8a is expressed as a heterodimer (CD8αβ) or homodimer (CD8αα). Most thymocytes and mature αβ T cells express CD8αβ, while γδ T cells, intestinal intraepithelial lymphocytes (IELs), and dendritic cells express CD8αα. CD8 binds to MHC class I and associates with tyrosine kinase p56lck, playing a role in T cell development and activation.

Each product contains:

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

Applications Reported: Flow cytometric analysis
Applications Tested: Tested on mouse splenocytes. Use ≤ 0.5 µg/test (in 100 µL). Adjust cell number empirically (10⁵–10⁸ cells/test). Optimize antibody titration for best performance.

Important Notes:

• NovaFluor dyes are incompatible with DNA intercalating viability dyes (e.g., propidium iodide, 7-AAD, DAPI).
• Use Invitrogen LIVE/DEAD Fixable Dead Cell stains for viability.
• CellBlox Plus Blocking Buffer (Cat. No. C001T06F01) is required for optimal staining.
  • Add 5 µL per sample to antibody cocktails before mixing with cells.
  • For single-color controls, use 5 µL per 100 µL of cell sample (10³–10⁸ cells).

Technology:
NovaFluor conjugates use Phiton™ technology with engineered fluorescent signatures minimizing spillover and spread.
Excitation: 552 nm; Emission: 700 nm; Laser: 561 nm (Yellow).

Strain-specific note:
NovaFluor Yellow 700 may non-specifically stain B cells in SJL mice. No non-specific staining observed in BALB/c or C57BL/6 strains.

Target Information

CD8 (Cluster of Differentiation 8) is a type I transmembrane glycoprotein of the immunoglobulin receptor family involved in signal transduction, T cell differentiation, and activation.

• Expressed as disulfide-linked CD8αβ heterodimers on T cells, functioning as co-receptors for MHC class I.
• Also expressed as CD8αα homodimers on subsets of T cells, NK cells, monocytes, and dendritic cells.
• Engagement with MHC-I/peptide complexes recruits Lck kinase, triggering lymphokine production, motility, and cytotoxic T lymphocyte (CTL) activation.
• Differentiation of naïve CD8⁺ T cells into CTLs is enhanced by IL-2, IL-12, and TGF-β1.

For Research Use Only. Not for use in diagnostic procedures.

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