Cyto-Last™ Buffer is specially formulated for the storage of cytokine producing cells. When used, staining of intracellular and/or extracellular targets for flow cytometric analysis can be delayed for up to two weeks. Cells should be stored at 4°C during this time. This unique buffer ensures cells maintain a background staining signal equal to that of freshly prepared cells, while also retaining high specific immunofluorescent staining against target antigens.Product Details
- Storage & Handling
- Store Cyto-Last™ Buffer between 2°C and 8°C.
ICFC - Quality tested
- Application Notes
1. Prepare target cells of interest and perform surface staining as described in BioLegend's Cell Surface Immunofluorescence Staining Protocol. (Note: staining with tandem-dye-conjugated antibodies (e.g., PE/Cy7, APC/Cy7, etc.) is not recommended as color compensation shifts may occur with long-term storage.)
2. Fix the cells with 0.5 mL/tube BioLegend's Fixation Buffer (Cat. No. 420801) at room temperature, in the dark, for 20 minutes.
3. Centrifuge at 350 x g for 5 minutes; discard supernatant.
4. Resuspend the cells in 0.5-1 mL/tube Cyto-Last™ Buffer, mix, cap the tubes, and then store at 4°C in the dark (Note: Cyto-Last™ Buffer can also be used for preserving cells in bulk at a cell concentration of 0.5-2.0 x 106 cells/mL.).
5. Take out the tubes at desired time points. Remove Cyto-Last™ Buffer by centrifugation, permeabilize the cells with BioLegend's Permeabilization Wash Buffer (Cat. No. 421002), and perform intracellular cytokine staining.
- Gene ID