CD4 Enrichment of Cultured Splenocytes Using MojoSort

January 31, 2020

Anatomy and Cell Biology
University of Illinois at Chicago
Graduate Student

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Company:

BioLegend

Product Name:

MojoSort Mouse CD4 T Cell Isolation Kit

Catalog Number:

480005

I've use the Biolegend MojoSort mouse CD4 T cell isolation kit with primary splenocytes I cultured in Th1-polarizing media. Following the manufacture protocol, I observed enrichment of CD4 from 12% in the starting culture to 95% in the post-MojoSort culture when I gate on Live cells. However, I also noticed a slight increase in dead cells after using MojoSort. So I would recommend subsequently using a cell separation media, like Lympholyte-M, to remove dead cells after using MojoSort.

Experimental Design and Results Summary

Applications

Cell Isolation/Separation

Sample

Primary mouse splenocytes

Primary Incubation

1:10, 15min, on ice

Blocking Agent

None

Secondary Incubation

1:10, 15min, on ice

Tertiary Incubation

No dilution. Magentic separation, 5min

Detection

Flow cytometer

Results Summary

Enrichment of CD4+ cells from 12% to 95%. No removal of dead cells or debris with MojoSort kit.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Fast and straight-forward protocol

The Bad

Proprietary antibody cocktail - unknown what cells are (and importantly are not) being marked for removal

The Bottom Line

The MojoSort Mouse CD4 T cell isolation kit works very well to enrich for CD4 cells in a primary splenocyte culture. When used in tandem with a dead cell separation medium like Lympholyte-M, high purity T cell cultures can be achieved.

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