Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Alexa FluorŪ 647 conjugated products with eFluorŪ 660 conjugated products.
Description
The streptavidin fluorochrome conjugates are commonly used in indirect staining protocols to detect biotinylated primary antibodies in flow cytometry. Streptavidin binds to biotin with high affinity.
Applications Reported
For research use only, not for diagnostic or therapeutic use. Streptavidin PerCP-eFluorŪ 710 has been reported for use in flow cytometric analysis.
Applications Tested
Streptavidin PerCP-eFluorŪ 710 has been tested by flow cytometric analysis of human peripheral blood cells. This can be used at less than or equal to 0.125 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 105 to 108 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
PerCP-eFluor® 710 can be used in place of PE-Cy5, PE-Cy5.5 or PerCP-Cy5.5. PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm). Please make sure that your instrument is capable of detecting this fluorochrome. For a filter configuration, we recommend using the 685 LP dichroic mirror and 710/40 band pass filter, however the 695/40 band pass filter is an acceptable alternative.
Our testing indicates that PerCP-eFluor® 710 conjugated antibodies are stable when stained samples are exposed to freshly prepared 2% formaldehyde overnight at 4°C, but please evaluate for alternative fixation protocols.
