Product: Ultra-LEAF™ Purified anti-mouse CD3ε

Catalog No.: 100339

Adam Coleman, Postdoctoral Fellow Oak Ridge Institute for Science and Education

Good positive control for CD4+ T cell proliferation (Ultra-LEAF™ Purified anti-mouse CD3ε)
Overall:	Product Quality:	Ease of Use:
Plates coated with anti-CD28/anti-CD3e cause CD4+ T cells to proliferate non-specifically.

Experimental Design

Application:	Cell proliferation
Cells used:	Mouse splenic CD4+ T cells
Brief Protocol:	Mash spleen through nylon mesh Purify CD4+ T cells by magnetic column purification Label cells with CFSE to track proliferation Coat 96 well round bottom plates with 4 ug/ml each Ultra-LEAF™ Purified anti-mouse CD28 and Ultra-LEAF™ Purified anti-mouse CD3e in 100 ul PBS at 37 degrees C for 2 hours Wash plates 3x with PBS Plate cells at 1e6 cells/ml in 200ul complete RPMI at 37 degrees C/5% CO2 for 72h Pellet cells Prepare cells for FACS by staining for viability (ie fixable Efluor 780 dye) and CD4+ cells (ie BV421 anti-mouse CD4) FACS analysis gated for live CD4+ cells
Results Summary:	CD4+ T cells proliferate well on surfaces coated with anti-CD28/anti-CD3e.
Additional Notes:

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Publication Status This data has not been published.

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