Flow Cytometry Staining Buffer is formulated and optimized for immunofluorescent staining of single-cell suspensions and subsequent dilution, suspension, washing, and storage of cells intended for flow cytometric acquisition and analysis. This product is supplemented with Bovine Serum Albumin (BSA) and the metabolic inhibitor sodium azide.
Intended Use:This product is designed for flow cytometry applications with non-conjugated or fluorochrome-conjugated antibodies. Refer to product specification sheets to obtain the recommended working antibody dilutions.
Surface Staining Protocol:1. Wash cells twice with 2ml of Flow Cytometry Staining Buffer, spinning the tube at 300 x g for 5 minutes
2. At this point, Fc receptor blocking reagents may be added to washed and counted cells. For antibodies directed to the Fc receptor follow the manufacturer’s recommendations. If using excess pre-immune IgG to block Fc receptors, use 1ug of IgG per 1x10e5 cells to be stained. The excess IgG does not need to be washed from the cells following the incubation period and can be carried into the staining reaction
3. For staining, transfer cells to a 5ml flow cytometry tube at a concentration between 1x10e6 and 1x10e7 cells per 100ul volume
4. Add antibody to the cells at the optimal concentration
5. Incubate the mixture for 30-45 minutes at 2-8°C
6. Following the incubation, remove any excess antibody by washing the cells twice in 2ml of Flow Cytometry Staining Buffer. Resuspend the final cell pellet in 200-40ul of Flow Cytometry Staining Buffer for flow cytometric analysis.
Stability and Storage:Upon receipt, Flow Cytometry Staining Buffer may be stored at 2-8° C for up to 6 months.
Precautions:
The safety and efficacy of this product in diagnostic or other clinical uses has not been established.
Contains 0.1% sodium azide. Sodium azide may react with lead and copper plumbing to form explosive metallic azides. Flush with large volumes of water during disposal.
