Figure 1. Staining performance and post-fixation stability. (A) Comparison of mouse splenocytes stained with CD8a antibody conjugated to either Super Bright 600 (red, Cat. No. 63-0081-82) or Brilliant Violet™ 605 conjugate (gray), at the same antibody concentration. (B) Mouse splenocytes were stained with CD45R antibody conjugated to Super Bright 600 (Cat. No. 63-0452-82) and either left unfixed (red), or were fixed in Invitrogen eBioscience IC Fixation Buffer (Cat. No. 00-8222-49) for 30 minutes (blue), 24 hours (orange), or 3 days (green).
Comparable to Brilliant Violet 605 conjugates
Invitrogen eBioscience Super Bright 600 antibody conjugates for flow cytometry provide:
- Brightness and resolution of positive and negative populations comparable to that of Brilliant Violet™ 605 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
Multiplexing with Super Bright 600 antibody conjugates
| 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 |
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Not for resale. Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.