Product: Alexa Fluor® 647 anti-human CD25

Catalog No.: 302617

Christopher Corkum, Research Assistant Memorial University of Newfoundland

Good Detection of CD25 on PHA-Stimulated PBMC (Alexa Fluor® 647)
Overall:	Product Quality:	Ease of Use:
CD25 surface expression was easily detected on lymphocytes stimulated in vitro with phytohemagglutinin.

Experimental Design

Application:	Flow cytometry
Cells used:	Human peripheral blood mononuclear cells
Brief Protocol:	Human PBMC were isolated from whole blood by standard Ficoll density gradient separation and cultured in AIM-V media supplemented with 10% fetal calf serum and 5ug/ml PHA for 72 hrs at a cell density of 1 x 10^6 per ml. After 72 hrs, cells were washed and approximately 1 x 10^6 Cells were incubated with the antibody for 1 hour on ice at 1:20 dilution in a total volume of 100ul PBS+2% FCS, 1mM EDTA, 0.1% sodium azide. Cells were washed twice with the above buffer and fixed with 4% PFA for 20 min on ice, followed by two additional washes.
Results Summary:	CD25 expression was bright on a significant proportion of the cells (black dots and histogram) and CD25+ cells were easily distinguished from CD25- cells and cells incubated with a matched isotype control (red dots and histogram).
Additional Notes:

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Publication Status This data has not been published.

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