Good Way to Purify CD4 or CD8 T Cells

Immunology
Duke University
Research Scientist

Overall

Quality of Results

Ease-of-Optimization

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Company:

Biolegend

Product Name:

MojoSort™ Streptavidin Nanobeads

Catalog Number:

480016

We use a mouse model to study CD4 and CD8 T cell signaling and activation. Pure CD4 and CD8 are needed for RT-PCR, Western blot, and all other functional assays. So it is necessary to perform the CD4 and CD8 purification. We are currently using this product and get very good purity and yield.

Experimental Design and Results Summary

Applications

Cell Isolation/Separation

Sample

Splenocytes

Primary Incubation

Biotin antibody cocktail

Blocking Agent

MojoSort Buffer

Secondary Incubation

Streptavidin Nanobeads

Tertiary Incubation

NA

Detection

NA

Results Summary

Unbound CD4 or CD8 purity is usually 95%.

DOI or PMID #

29386256

Additional Notes

The purification steps are : 1. Resuspend the cells in MojoSort™ Buffer in a 5mL (12 x 75 mm) polypropylene tube, 107 cells/100µL. 2. Add 10l of antibody cocktail to 100l cell suspension, incubate on ice for 15 minutes. 3. Resuspend Streptavidin Nanobeads by vortexing. Add 10l beads to the tube, mix well, and incubate on ice for 15 minutes. 4. Add 2ml MojoSort™ Buffer to the tube and mix well. 5. Place the tube in the magnet for 5 minutes. 6. Collect the liquid in a new tube while the tube is in the magnet. The cells in the liquid are untouched cells (negative selection). If you are interested in the cells that are bound to the Nanobeads (positive selection), remove the tube from the magnet and collect the cells.

Image Gallery

Summary

The Good

Good purity and reasonable yield

The Bad

NA

The Bottom Line

Easy method and reproducible

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