Naive CD4+ T Cell Isolation Kit - Our Gold Standard

February 05, 2019

University of Michigan
Pathology
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Miltenyi Biotec

Product Name:

Naive CD4+ T Cell Isolation Kit, mouse

Catalog Number:

130-104-453

For a CD4 T cell lab, we use a lot of these kits. We've tried others, but this has always come out on top for quality. The bottom line: For labs that do extensive T cell work, this is strongly recommended for large number and high quality purification.

Experimental Design and Results Summary

Application

Ex vivo naive CD4+ T cell isolation

Starting Material

Mouse splenocytes

Protocol Overview

Single cell suspension of mouse splenocytes was used. Samples were RBC lyses with ACK buffer, then washed with MACS (PBS+BSA+EDTA) and incubated with antibody cocktail. Following 15 minute incubation (4C), cells were washed and incubated with anti-biotin microbeads for 10 minutes, then processed with an autoMACS Pro using the "depletes" function. Expected yield is 3-5 million naive T per 100 million input cells.

Tips

RBC lysis can be used to lower the antibody volume required (approximately 1/3 of the recommended amount even). Peripheral LN, such as mesenteric and inguinal, can be used in addition to spleen.

Results Summary

Purity and yield will vary inversely, based on the volume of antibody used per cell number. For high purity, more antibody is required, but will result in decreased yield. Quality of cells (e.g. less debris, higher viability) is better than simpler magnetic columns (e.g. MojoSort) in our hands.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Quality, reproducible purification.

The Bad

Cost is higher than others, requires autoMACS or similar instrument.

The Bottom Line

For labs that do extensive T cell work, this is strongly recommended for large number and high quality purification. Entry cost is the biggest issue, and kit and column cost add up over time, but the results are good.

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