APC Fire 750

Multicolor Compatibility of APC/Fire™ 750

PMA+Ionomycin (6 hours, in the presence of Monensin) stimulated human PBMCs were stained with anti-CD4 Alexa Fluor® 647 and then treated with Fixation Buffer followed by permeabilization with 1X Intracellular Staining Permeabilization Wash Buffer and stained with anti-IFN-γ APC/Fire™ 750 for 30 min.

Brightness Comparison

Human whole blood was stained for 20 min with anti-CD3 conjugates of APC/Fire™ 750, APC/Cy7, APC-H7, or APC-eFluor® 780 at each manufacturer's recommended optimal dilution, followed by RBC lysis and wash steps. Histograms were gated on lymphocyte populations based on forward and side scatter.

Low Background Binding to Monocytes 

Human whole blood was stained with anti-CD3 conjugates of APC/Fire™ 750 or APC/Cy7. Histograms were gated on monocyte populations.

APC/Fire™ 750 Stability

Human whole blood was stained for 20 min with anti-CD3 conjugates of APC/Fire™ 750, APC/Cy7, APC-H7, or APC-eFluor® 780 at each manufacturer's recommended optimal dilution, followed by RBC lysis and wash steps. Cells were then treated with either PBS control, 1% paraformaldehyde (PFA) in PBS, 4% PFA followed by 0.1% saponin, BioLegend's True-Nuclear™ Fix/Perm Buffer Set, BD's Transcription Factor Buffer Set, or eBioscience's FoxP3/Transcription Factor Staining Buffer Set prior to analysis. Lymphocyte populations were gated on based on optimal forward and side scatter.

Heat Stability 

A vial of APC/Cy7 or APC/Fire™ 750 was stored in the dark at either 4°C or 37°C for 81 days. At certain intervals, an aliquot of reagent was used to stain Veri-Cells™, a lyophilized human PBMC product that effectively removes donor dependent variation in staining. Cells were stained for 15 min at room temp in cell staining buffer.