Purifying untouched lymphocytes the easy way:no kits, no ice

•Do you

•Routinely purify mouse lymphocytes (CD4, CD8 or B)?

•Use Wild Type, not T cell transgenic, mice?

•Process several samples individually (cell sorting not feasible)?

•Prefer untouched purified cells (cell sorting inconvenient)?

•Desire APC-free purified lymphocytes?

•Desire good recovery (>50%) of the starting population?

•Find commercial kits disappointing?

•Then this protocol is for you

Pure untouched CD4+CD3+ T cells using a prototype antibody cocktail and negative selection

Excellent purity

Very high recovery

CD4+CD3e+ percentage

Pre Average

Post

Average

%pre-depletion CD4+CD3e+

cells recoveredAverag

e

33.9 31.2 98.

2 97.7 52 74.3

25.0

96.5 70

20.3

96.3 95

33.8

98.2 88

32.4

98.0 77

41.7

99.0 64

Compared to commercial CD4 enrichment kits, custom CD4 and CD3 enrichment kits are better at depleting APCs

Commercial kits are poor

Custom kits are excellent

Want more details?

•Check out and download my article free from WILEY InterScience:

http://onlinelibrary.wiley.com/doi/10.1111/j.1365-3083.2007.02066.x/pdf

•Citation Details:

Kamala T.

An optimized immunomagnetic bead-based negative selection protocol for CD4 T-cell isolation from mouse lymph nodes and spleen.

Scand J Immunol. 2008 Mar;67(3):285-94.