Great Kit for Purifying Untouched CD3+ T cells from PBMCs

Stanford University
Pediatrics
Postdoctoral Research Fellow

Overall

Performance

Ease-of-Optimization

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Company:

Miltenyi Biotec

Product Name:

Pan T Cell Isolation Kit, human

Catalog Number:

130-096-535

Love this kit. It's easy, fast, and efficient. Routinely get >95% purity of CD3+ cells from buffy coat-derived PBMCs. I can highly recommend this kit.

Experimental Design and Results Summary

Application

T cell purification

Starting Material

PBMCs from human buffy coats

Protocol Overview

Ficoll-based PBMC extraction.Red blood cell lysis using Ammonium Chloride Solution.Spin down desired number of starting PBMCs and resuspend in 400ul MACS buffer.Add 100ul Pan T Cell Biotin-Antibody Cocktail and mix. Incubate 5min at 4°C.Add 300ul MACS buffer and 200ul Pan T Cell MicroBead Cocktail and mix.Incubate 10min at 4°C.Set up column on MACS magnet and equilibrate with 3ml MACS buffer.Add cells to column and collect flow-through (negative selection kit).Wash column with 5ml MACS buffer and collect flow-through.

Tips

Follow the protocol and you'll be fine

Results Summary

From 100 million PBMCs, yields typically range from 20-40 million cells with viabilities and purities around 90% or higher.

Features Summary

Isolation of a pure and untouched T cell population

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Easy, fast, and efficient

The Bad

A bit on the expensive side

The Bottom Line

Great product. Would highly recommend.

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