BD Horizon Brilliant™ Stain Buffer

1 BD Horizon Brilliant Stain Buffer Optimizing Experiments Using BD Horizon Brilliant Polymer Conjugates Features Mitigates Non-specific Interactions BD Horizon Brilliant Violet (BV), Ultraviolet (BUV), and Blue Improves staining quality when multiple BD Horizon (BB) polymer conjugates have expanded the number of bright Brilliant polymer conjugates are used in the same fluorochromes available for multicolor flow cytometry. As with experiment all fluorochromes, building multicolor panels with these reagents Allows pre-mixing of cocktails containing multiple requires optimization. conjugates for at least 24 hours before use While using multiple BD Horizon Brilliant reagents in the same Provided free of charge with all BD Horizon Brilliant experiment, we observed several artifacts that appeared to conjugates be related to non-specific interactions between the polymer dyes themselves.

2 In some instances populations appear to be undercompensated in multicolor staining, while single color 4h compensation controls appear to be properly compensated. In 0h 4 C other instances the negative population can appear multivariate No Buffer No Buffer rather than as a single, discrete population. The effects of interactions can be seen in reagents from BD and from other 105. 105. companies BD Horizon Brilliant Stain Buffer , when added to samples 104. 104. CD8 BV650. CD8 BV650. prior to addition of antibodies, can mitigate these interactions, improving staining quality regardless of the source of reagents. 103. 103. The solution also can be used to stabilize antibody cocktails containing BD Horizon Brilliant polymer conjugates for at least 102. 102. 24 hours. 0. 0. -221. -221. M. -180.

3 0 102 103 104 105 M. -180. 0 102 103 104 105 Applicable to Many Experiments CD4 BV421 CD4 BV421 The various interactions appear to be dependent upon several factors, including reagent combination, staining volume, sample 4h 24 h type, and variation between samples. Regardless of the cause, 4 C 4 C. With Buffer With Buffer the interactions can be mitigated with the addition of Brilliant Stain Buffer prior to the addition of antibodies to the samples. 105. 105. Since there are so many variables that can contribute to the interactions, Brilliant Stain Buffer might not be required in all 104. 104. experiments. However, we recommend using Brilliant Stain Buffer CD8 BV650. CD8 BV650. any time more than one BD Horizon Brilliant polymer conjugate is 103. 103. used in the same experiment. As with any new protocol, we highly recommend that 102.

4 102. investigators perform side-by-side experiments to determine 0. 0. if their staining can benefit from the addition of Brilliant Stain -221. -221. M 0 102 103 104 105 M 0 102 103 104 105. -180 -180 Buffer . To facilitate this, BD provides a complimentary vial of CD4 BV421 CD4 BV421. Brilliant Stain Buffer with every BD Horizon Brilliant antibody Figure 1. Brilliant Stain Buffer improves the performance of BV dyes in purchase. Customers not purchasing BD antibodies can purchase cocktails. the Buffer separately (Cat. No. 563794). Antibody cocktails were prepared in the presence or absence of Brilliant Stain Buffer for the designated period of time at 4 C protected from light. Lysed human whole blood samples were subsequently stained with the cocktails. In the absence of Brilliant Stain Buffer , the CD4+ population shows non-specific interaction.

5 Addition of Brilliant Stain Buffer restored the populations to their expected locations. Visit for more information. For Research Use Only. Not for use in diagnostic or therapeutic procedures. BD Horizon Brilliant Stain Buffer BD Other Company A. 105. 105. 104. 104. BV711. BV711. Without Brilliant 103. 103. Stain Buffer 0. 0. -640. -640. M 0 103 104 105 M 0 103 104 105. -472 -472. BV605 BV605. 105. 105. 104. 104. BV711. BV711. With Brilliant 103. 103. Stain Buffer CD8 BV711. 0. 0. -640. -640. M 0 103 104 105 M 0 103 104 105. -472 -472. BV605 BV605. CD19 BV605. BD Other Company B. 105. 105. 104. 104. BV711. BV711. Without Brilliant 103. 103. Stain Buffer 0. 0. -472. -472. M 0 103 104 105 M 0 103 104 105. -472 -472. BV605 BV605. 105. 105. 104. 104. BV711. BV711. With Brilliant 103. 103. Stain Buffer CD8 BV711.

6 0. 0. -472. -472. M 0 103 104 105 M 0 103 104 105. -472 -472. BV605 BV605. CD19 BV605. Figure 2. Enhanced performance of BV reagents. CD19 BV605, CD8 BV711, and CD4 BV421 from BD or from another company were used to Stain (A) peripheral blood mononuclear cells or (B) lysed whole blood. In the absence of Brilliant Stain Buffer , these mutually exclusive populations can appear under-compensated. Note the multivariate negative population, where the red population corresponds to the CD8/CD19. double negative population and the green population corresponds to the CD4+ population. In both instances the addition of Brilliant Stain Buffer eliminates the issue and staining returns to the expected pattern. Ordering Information Description Size BD Horizon Brilliant Stain Buffer 100 Tests 563794. For Research Use Only.

7 Not for use in diagnostic or therapeutic procedures. BD, BD Logo and all other trademarks are property of Becton, Dickinson and Company. 2014 BD BD Biosciences 23-15643-02