What is BD APC-H7?
BD APC-H7 is a newly designed APC-cyanine tandem conjugate that is more stable and less affected by light, temperature, and formaldehyde-based fixatives than other APC-cyanine tandems. H7 is an analog of Cy7, a cyanine-based dye.
Why did BD develop BD APC-H7 tandem conjugates?
The increased spillover into the APC channel due to APC-cyanine tandem degradation has been a challenge for some customers needing to keep experimental samples exposed to room temperatures or paraformaldehyde-based fixatives for any extended period of time. This spillover reduced sensitivity in the APC channel and resulted in insufficient resolution and increased experimental variability in this and other nearby channels. To address this problem, BD Biosciences engineered an APC-cyanine with enhanced stability and performance in these situations.
Is APC-H7 a superior dye to APC-Alexa Fluor® 750?
In our tests, APC-H7 outperformed APC-Alexa Fluor® 750 in terms of stability (as measured by % spillover) when exposed to light, temperature, and paraformaldehyde-based fixatives. In fact, APC-H7 exibited significantly less initial (out of the bottle) spillover than APC-Alexa Fluor® 750 conjugates.
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.
What are the limitations of APC-H7?
APC-H7 offers greater stability when exposed to light, room temperature, and paraformaldehyde-based fixatives when compared to other APC-cyanine tandem dyes, including APC-Cy7 and APC-Alexa Fluor® 750. However, the stain index (brightness of staining over background) of freshly stained samples is slightly less than APC-Cy7. This reduced staining intensity is most evident to customers who are not currently experiencing increased spillover into the APC channel as a result of tandem degradation. For those customers who do experience reduced sensitivity and resolution because of APC-cyanine tandem degradation, using the more stable APC-H7 will offer improved performance, even with a reduced staining intensity.
Will BD still support its APC-Cy7 products?
Yes. A large majority of our research customers report no trouble when using APC-Cy7 conjugates in their multicolor experiments. With proper precautions and controls and minimized exposure to light, elevated temperature, and fixatives, APC-Cy7 conjugates can continue to be used in multicolor experiments without worry by many who want a proven, quality flow cytometry reagent.
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