Ignite your flow cytometry research with APC/Fire™ 750.

About APC/Fire™ 750

BioLegend is excited to announce the release of our newest tandem APC/Fire™ 750.

APC/Fire™ 750 provides a more temperature and photo-stable alternative to APC/Cy7. APC/Fire™ 750 also has lower compensation requirements than APC/Cy7 conjugates while maintaining an equal level of brightness. Additionally, APC/Fire™ 750 has minimal non-specific binding to monocytes, as has been observed with APC/Cy7. Lastly, APC/Fire™ 750 conjugates are consistently brighter than APC-H7 in all conjugates tested.

As with all BioLegend products, antibody conjugates are provided at the best value for researchers along with expert technical support and a 100% satisfaction guarantee.

 

 

Excellent Resolution and Minimal Spillover into APC

image

PMA+Ionomycin (6 hours, in the presence of Monensin) stimulated human PBMCs were stained with CD4 Alexa Fluor® 647 and then treated Fixation Buffer (Cat 420801) followed by permeabilization with 1X Intracellular Staining Permeabilization Wash Buffer (Cat 421002) and stained with IFN-γ APC/Fire™ 750 for 30 min.

Improved Stability

image

The 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 (cat. # 425002). Cells were stained for 15 min at room temp in cell staining buffer.

Comparison with APC-H7 and Other Equivalents

image

Human whole blood was stained for 20 min with CD3 (SK7) 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

image

Human whole blood was stained for 20 min with CD3 (SK7) 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.

Comparison Stability in Fixative Solutions

image

Human whole blood was stained for 20 min with CD3 (SK7) 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.

Comparison Summary Table

  APC/Fire™ 750 APC/Cy7 APC-H7 APC-eFluor® 780
Brightness ++ ++ + ++
Stability to Fixatives ++ - ++ -
Photostability ++ - ++ NT
Compensation into APC ++ + ++ +
Low Monocyte binding ++ - ++ +
Long-term storage stability ++ - + NT
Price per test/µg + ++ - +

 

Legend: (-) = poor, (+) = moderate, (++) = best, NT = not tested.

 

For a complete listing of the trademarks and patents referenced on this page, view our Trademarks page.

Login/Register
Forgot your password? Reset Password
Request an Account