Product: Alexa Fluor® 488 anti-mouse/human CD45R/B220

Catalog No.: 103225

Javier Carrero, Assistant Professor Washington University School of Medicine

Alexa Fluor® 488, Brighter Alternative to B220-FITC
Overall:	Product Quality:	Ease of Use:
This is the best anti-B220 reagent we have tested for the 488 nM channel.

Experimental Design

Application:	Flow Cytometry
Cells used:	lymph nodes and tissue resident macrophages
Brief Protocol:	1) Isolated cells and made single cell suspension using collagenase/DNAse I. 2) Washed cells with wash buffer (PBS, 1% bovine serum albumin). 3) Blocked Fc receptors with anti-CD16/32 (clone 2.4G2) at 500ng/mL in wash buffer at a final volume of 100 microliters per million cells. 4) Washed cells with wash buffer. 5) Resuspended cells in 100 microliters and added 0.5 microliters of antibody (1:200) per million cells. Mixed well and incubated at 4 degrees centigrade for 30 minutes. 6) Washed cells twice with wash buffer. 7) Added 2% paraformaldehyde in PBS to fix the cells. 8) Washed with wash buffer once. 9) Kept at 4 degrees centigrade until ready to analyzed (up to 24 hours). All steps done in a 96 well V-bottom plate.
Results Summary:	We obtain a 1.5 to 2 log shift from unstained or isotype controls on a BD FACSCanto II instrument.
Additional Notes:	None.

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Publication Status This data has not been published.

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