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Super Bright 645 Antibody Conjugates
Comparable to Brilliant Violet 650 conjugates
Invitrogen eBioscience Super Bright 645 antibody conjugates for flow cytometry provide:
- Less spill over into other violet channels than Brilliant Violet™ 650 conjugates
- Options in marker and clone selection for violet laser excitable antibody conjugates
- Compatibility with standard intracellular buffers, viability stains, compensation beads, and other antibodies
Figure 1. Fluorescence intensity comparison of Super Bright 645 conjugates and Brilliant Violet™ 650 conjugates.(A) Mouse splenocytes stained with anti-CD8a conjugated to Super Bright 645 (red, Cat. No. 64-0081-82) or Brilliant Violet™ 650 conjugate (gray), at the same concentration of antibody. (B) Human peripheral blood cells stained with anti-CD8a conjugated to Super Bright 645 (red, Cat. No. 64-0088-42) or Brilliant Violet™ 650 conjugate (gray), using the same concentration of antibody.
| Multiplexing compatibility | Buffer | Multiplexing considerations |
|---|---|---|
| Multiplexing with 1 Super Bright antibody conjugate | Standard buffers applicable | No special buffer required when only one Super Bright antibody conjugate is used in a panel |
| Multiplexing with 2 or more Super Bright antibody conjugates | Super Bright Staining Buffer | Special staining buffer is required prior to addition of Super Bright conjugates to reduce non-specific dye-dye interactions |
| Multiplexing with 1 or more Brilliant Violet™ antibody conjugates | Super Bright Staining Buffer | Special staining buffer is required to reduce non-specific dye-dye interactions. Use of the Super Bright Staining Buffer is recommended, but the similar Brilliant Stain Buffer can be substituted |
Figure 2. Multiplex flow cytometry experiment. Normal human peripheral blood cells were diluted in Super Bright Staining Buffer, then surface stained with the indicated reagents. Viable cells (determined using a fixable viability stain) were used for analysis to discriminate various T cell subsets.
| Viability stain options | Product | Multiplexing considerations |
|---|---|---|
| Fixable | LIVE/DEAD fixable dead cell stain kits | No compatibility concerns |
| Non-fixable | SYTOX non-fixable dead cell stains Ready Flow Ready-to-use viability reagents | No compatibility concerns Compatible with all Ready Flow reagents for viability |
| Product | Multiplexing considerations |
|---|---|
| UltraComp eBeads microspheres | UltraComp eBeads microspheres are compatible but OneComp eBeads are not compatible with violet lasers; the AbC Total Antibody Compensation Bead Kit is also compatible with the Super Bright antibody conjugates. |
| Staining Target | Product | Multiplexing considerations |
|---|---|---|
| Cytosolic staining (cytokines) | Intracellular Fixation & Permeabilization Buffer Set | No compatibility concerns |
| Nuclear staining (transcription factors) | Foxp3/Transcription Factor Staining Buffer Set | No compatibility concerns |