Human Multipotent Mesenchymal Stromal Cell Multi-Color Flow Cytometry Kit

Product Description
This kit contains four conjugated antibodies (and corresponding isotype controls) that can be used for single-step staining of human multipotent mesenchymal stromal cells (hMSCs) (1 - 4):

Positive Markers
• CD105-PerCP (Clone 166707; mouse IgG1)
• CD146-CFS (Clone 128018; mouse IgG1)
• CD90-APC (Clone Thy-1A1; mouse IgG2A)

Negative Marker
• CD45-PE (Clone 2D1; mouse IgG1)

This kit also contains Staining Buffer (100 mL).

Intended Use
This product is designed for the flow cytometric analysis of hMSCs using four fluorochrome-conjugated antibodies.

Storage
Store at 2 - 8° C in the dark. Use within 6 months of receipt.

Precaution
The Staining Buffer 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.

Surface Staining Protocol
1. Cell samples should be washed with 2 mL of Staining Buffer, spinning the tube at 300 x g for 5 minutes.
2. Washed cells should be counted and then Fc receptor blocking reagents may be added. If using excess pre- immune IgG to block Fc receptor, use 1 μg of IgG per 1 x 105 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. Transfer a small volume (about 100 μL) of the Fc receptor-blocked cells (about 1 x 106
cells) into a 5 mL Flow Cytometry tube.
4. Add 10 μL of each antibody or each corresponding isotype control antibody to the cells.
5. Incubate the mixture for 30 - 45 minutes at 2 - 8° C in the dark.
6. Following the incubation, remove any excess antibody by washing the cells with 2 mL of Staining Buffer. The final cell pellet is resuspended in 200 - 400 μL of Staining Buffer for flow cytometric analysis.