Super Bright 645 versus 650 conjugates

Comparable to Brilliant Violet 650 conjugates

Invitrogen eBioscience Super Bright 645 antibody conjugates for flow cytometry provide:

Data comparison

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.

Antibodies
Multiplexing compatibilityBufferMultiplexing considerations
Multiplexing with 1 Super Bright antibody conjugateStandard buffers applicableNo special buffer required when only one Super Bright antibody conjugate is used in a panel
Multiplexing with 2 or more Super Bright antibody conjugatesSuper Bright Staining BufferSpecial 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 conjugatesSuper Bright Staining BufferSpecial 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 stains
Viability stain optionsProductMultiplexing considerations
FixableLIVE/DEAD fixable dead cell stain kitsNo compatibility concerns
Non-fixableSYTOX non-fixable dead cell stains

Ready Flow Ready-to-use viability reagents		No compatibility concerns

Compatible with all Ready Flow reagents for viability
Compensation beads
ProductMultiplexing considerations
UltraComp eBeads microspheresUltraComp 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.
Intracellular staining buffers
Staining TargetProductMultiplexing considerations
Cytosolic staining
(cytokines)		Intracellular Fixation & Permeabilization Buffer SetNo compatibility concerns
Nuclear staining
(transcription factors) 		Foxp3/Transcription Factor Staining Buffer SetNo compatibility concerns

Resources

Support

Not for resale. Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.