Biotin Cell Separation Kit

Albert Einstein College of Medicine
Microbiology & Immunology
PhD Candidate

Overall

Quality of Results

Ease-of-Optimization

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Company:

Stemcell Technologies

Product Name:

EasySep™ Biotin Positive Selection Kit

Catalog Number:

18559

We use this kit to purify cells that have been successfully transduced with a lentivirus expressing a truncated mouse reporter protein. It can be used for any kind of cell selection when using the appropriate biotynilated antibody.

Experimental Design and Results Summary

Application

Cell separation

Starting Material

Transduced human CD8+ T cells

Protocol Overview

Cells are resuspended at 10^8 cells/ml. Add biotynilated antibody at optimal concentrations (I use 1:100), incubate for 15min at room temperature (RT). Without washing add biotin selection coctain at 1:10, incubate 15min at RT. Mix magnetic particles and add 50ul/ml., incubate 10min at RT. Bring volume up to 5ml, place in magnet for 5min. Wash 3x with 5ml of buffer. Resuspend positive cells as desired.

Tips

Follow the protocol.

Results Summary

We get very high purity of cells expressing our reporter protein (>90%). Cells are viable and not affected by the purification.

Additional Notes

None

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Summary

The Good

High purity, easy protocol

The Bad

None

The Bottom Line

Good kit to use for purification of cells based on expression of surface markers/reporters.

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