Total CD4+ T Isolation Kit

University of Michigan
Pathology
Graduate Student

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Quality of Results

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Company:

Miltenyi Biotec

Product Name:

CD4+ T Cell Isolation Kit, mouse

Catalog Number:

130-104-454

For general T helper cell isolation, the yields from this are quite good. Useful for both lymphoid and non-lymphoid tissues.

Experimental Design and Results Summary

Application

Ex vivo isolation of CD4+ T cells.

Starting Material

Splenocytes or other tissue single cell suspensions

Protocol Overview

Cells are RBC lysed (depending on tissue) and resuspended in MACS (PBS+BSA+EDTA) and incubated in antibody cocktail for 15 minutes on ice. Cells are then washed and incubated with anti-biotin microbeads for 10 minutes, then processed with a autoMACS Pro using "depletes" function.

Tips

RBC lysis lowers required antibody amounts. Tissue type affects yield and quality.

Results Summary

We commonly use for isolation of T helper cells from lymphoid and non-lymphoid tissues for either culture or RNA extraction. Quality of cells is good. Typical yield is 15-20 million T helper cells per 100 million splenocytes (RBC lysed).

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

High yield

The Bad

Cost is fairly high. Requires autoMACS or other magnetic sorter.

The Bottom Line

A good product if you already have access to an autoMACS instrument, but if you don't commonly do this type of work, a cheaper alternative could be considered.

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