More reliable and reproducible staining patterns from the most stable APC-cyanine tandem dye on the market
BD™ APC-H7, a new APC-cyanine tandem dye, gives researchers more reliable and reproducible staining patterns from this family of dyes. Until now, emission at the donor fluorochrome wavelength, caused by dye degradation and spillover, has been a challenge in the use of tandem dyes such as APC-cyanine dyes.
BD APC-H7 is engineered to minimize spillover to the APC channel. In addition, the new BD APC-H7 is more stable and less affected by light, temperature, and formaldehyde-based fixatives, compared to other APC-cyanine tandem dyes such as APC-Cy™7 and APC-Alexa Fluor® 750. This reduces loss of fluorescence resonance energy transfer (FRET) between the donor and acceptor fluorophores to maintain integrity of signals from both channels.
More flexibility and greater ease of use for multicolor flow cytometry
Degradation inherent in tandem dyes leads to emission in the APC channel, which in turn leads to false positive events and reduced sensitivity. BD APC-H7 improves stability in ambient light, making it easier to handle and use. Experiments can be run outside darkroom conditions with more relaxed time constraints. Spillover is a common issue with multicolor applications, and high spillover can result in loss of resolution sensitivity. Reduced spillover in BD APC-H7 allows for greater reliability and reproducibility of results.
Conjugates of BD APC-H7 are available for many important cell surface markers such as CD3, CD4, CD8, CD14, CD19, CD20, CD45, and anti-HLA–DR, allowing BD APC-H7 to be used for a range of applications. This degree of choice, coupled with increased stability, enables greater flexibility in multicolor experiment design.
BD APC-H7 demonstrated stability and reduced spillover. The experiments shown here compare the stability, as measured by % spillover, of whole blood stained with BD APC-H7 and APC-Alexa Fluor® 750 conjugated antibodies. Stained samples were stored in 1% PFA at room temperature under ambient light.
BD APC-H7 conjugated monoclonal antibody reagents
BD APC-H7 is a tandem conjugate and an analog of APC-Cy7 with the same spectral properties. It is engineered for greater stability and less spillover in the APC channel. It has an absorption maximum of approximately 650 nm. When excited by light from a red laser, the APC fluorochrome can transfer energy to the cyanine dye, which then emits at a longer wavelength. The resulting fluorescent emission maximum is approximately 767 nm.
BD APC-H7 vs. APC-Cy7: Comparison of staining stability (in BD Stabilizing Fixative, under light at RT). Whole blood stained with CD4 APC-H7, CD8 APC-H7, CD4 APC-Cy7, or CD8 APC-Cy7. Stained samples were stored in BD Stabilizing Fixative at room temperature under ambient light for 48 hours.
BD APC-H7 vs. APC-Cy7: Comparison of staining stability (in 1% PFA, under light at RT). Whole blood stained with CD4 APC-H7, CD8 APC-H7, CD4 APC-Cy7, or CD8 APC-Cy7. Stained samples were stored in 1% PFA at room temperature under ambient light for 48 hours.
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