This gentle and easy-to-use kit allows you to isolate highly pure and functional mouse Treg cells from mouse lymph nodes or spleen samples. Your cells are not exposed to the stress of being passed through a column. The isolated CD4+CD25+Foxp3+ Treg cells are bead-free and flow-compatible. The cells can be readily expanded and retain their functional suppressive capacity. You can expect a purity level of >90% Treg cells. Over 88% of the isolated CD25+ cells express Foxp3. You will save time and money, and get increased confidence in your results.

Methodology:
Negative isolation of CD4+ T cells, followed by positive isolation of CD25+ regulatory T cells. First, an antibody mix is added to the sample and the Ab will bind the non-CD4+ T cells. Mouse Depletion Dynabeads™ are then added and will bind to the Ab-cell complex during a short incubation. Bead-bound cells are separated on a magnet and discarded. The remaining untouched mouse CD4+ T-cells are then labeled with FlowComp™ CD25 Antibody and positively isolated using FlowComp™ Dynabeads™. A short incubation with FlowComp™ Release Buffer then releases the isolated mouse regulatory T-cells from the Dynabeads™. The isolation method is easy to use and to scale up.

Advantages:

High purity of CD4+CD25+Foxp3+ Treg cells.

• No risk of your results being affected by the isolation method
• Gentle tube-based method - no columns needed
• High purity, recovery, and viability
• High CD25+Foxp3+ expression.
• Bead-free cells - compatible with flow cytometry analysis
• Treg cells are readily expanded and retain their functional suppressive capacity.

Starting sample:
spleen or lymp node cells