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As a global market leader with numerous subsidiaries and distributors, Miltenyi Biotec is committed to providing our customers around the world with the highest quality products. In addition to direct selling in more than 20 countries in North America, Europe and Asia/Pacific, Miltenyi Biotec also provides support for our customers through an extensive distributor network covering dozens of additional countries.
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This application protocol describes use of the Treg Expansion Kit, human, which is designed to efficiently expand Treg cells that maintain FoxP3 expression after isolation with the CD4+CD25+CD127dim/– Regulatory T Cell Isolation Kit II, human. It includes a complete protocol for your Treg cell workflow, describing human Treg cell isolation from peripheral blood mononuclear cells (PBMCs), their in vitro expansion, and subsequent flow cytometry analysis.
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Notes:
Example:
For 1 sample with 1×106 Treg cells: prepare 1 mL of fixation/permeabilization solution and 4 mL of 1× Permeabilization Buffer.
The expansion medium can be stored for up to 10 days under sterile conditions at 4 °C.
500 mL TexMACS™ Medium
+ 500 IU/mL Human IL-2 IS, premium grade
+ 5% human AB Serum
+ 0.01 mM β-ME
+ 1% PenStrep
+ (Optional) 100 nmol/L rapamycin
▲ Note: Rapamycin has been shown to increase the frequency of FoxP3+ Treg cells during expansion (Battaglia, M. et al. (2005) Blood 105: 4743–4748).
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