Product: Biotin Annexin V

Catalog No.: 640904

Jordan Chinai, Graduate Student Albert Einstein College of Medicine

Biotin Annexin V - Magnetic Dead Cells/Debris Removal
Overall:	Product Quality:	Ease of Use:
Worked great.

Experimental Design

Application:	Depletion of Dead Cells and Debris from Tumor Digestion
Cells used:	Tumor
Brief Protocol:	~300-500 mg of tumor was enzymatically digested. Cells were washed and resuspended in Annexin binding buffer. Cells were washed once more then resuspending in 200 uL of 1:50 dilution of Annexin-biotin for 20 minutes at room temperature. Cells were washed once with binding buffer then incubated with magnetic streptavidin beads at room temperature for 20 minutes Cell suspension was then incubated on magnet and the negative fraction was retained.
Results Summary:	My tumor digestions contained a lot of dead cells and debris that interfere with downstream stimulation and other applications. This was highly efficient for removing dead cells and debris.
Additional Notes:	Results varied slightly by tumor but typically 70-95% of dead/cells debris were removed as assessed by trypan blue staining on a hemacytometer. Subsequent staining we do results in more dead cells, but representative FSC/SSC is shown in image. Really nice purification of tumor infiltrating lymphocytes.

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