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Super Bright 436 Antibody Conjugates
Comparable to Brilliant Violet 421 conjugates
Invitrogen eBioscience Super Bright 436 antibody conjugates for flow cytometry provide:
- Less background than Brilliant Violet™ 421 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 436 antibody conjugates
Figure 1. Super Bright 436 dye performance comparison. Mouse bone marrow cells were stained with Anti-Ly-6A/E (clone D7) APC and either Anti-CD117 (clone 2B8) conjugated to (left panel) Super Bright 436 dye, (middle panel) eFluor 450 dye or (right panel) Brilliant Violet™ 421 dye.
Figure 2. Fluorescence intensity for Super Bright 436 dye conjugates compared to Brilliant Violet™ 421 conjugates. (A) Human peripheral blood cells were stained with CD19 antibody conjugated to either Super Bright 436 (purple, Cat. No. 62-0199-42), eFluor 450 dye (blue, Cat. No.48-0199-42), or Brilliant Violet™ 421 dye (orange). (B) Human peripheral blood cells were stained withCD27 antibody conjugated to either Super Bright 436 (purple, Cat. No. 62-0279-42) or Brilliant Violet™421 dye (orange).
| 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 3. 10-color ILC2 subset panel. Normal human PBMCs were surface-stained in the presence of Super Bright Staining Buffer (Cat. No. SB-4400-42) at optimal concentrations for the indicated surface markers.(A) Gated on live cells, this plot shows the lineage markers used as a FITC dump channel (CD3 (clone UCHT1), CD4 (clone SK3), CD8a (clone RPA-T8), CD11b (clone ICRF44), and CD19 (clone HIB19) vs. CD127 (IL-7RA) (clone eBioRDR5) Super Bright 436 (Cat. No. 62-1278-42) stained cells. Since ILC2 subsets are negative for all five of these markers, all CD127 and lineage-positive cells can be eliminated from further analysis. (B-G) Gating strategy is shown for all lineage targets to highlight the ILC2 population. (H, I) The CD294 (CRTH2) population is identified.
| Viability stain options | Product | Multiplexing considerations |
|---|---|---|
| Fixable | LIVE/DEAD fixable dead cell stain kits | Not compatible with LIVE/DEAD Fixable Violet Dead Cell Stain or Fixable Viability Dye eFluor 450 |
| Non-fixable | SYTOX non-fixable dead cell stains Ready Flow Ready-to-use viability reagents | Not compatible with SYTOX blue stain 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 |