Abcam
Anti-CD14 antibody [MEM-15] (FITC)
ab28061
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We isolated peripheral blood mononuclear cells from fresh human blood using Ficoll Paque Plus protocol and stained them with anti CD14-FITC antibody (MEM-15) Abcam catalog # ab28061 to identify monocyte population. The fixed and anti CD14-stained cells were plated on Concanavalin A treated 96 well imaging plate. Images were acquired using Highthroughput ImageXpress wide field microscope.
Immunofluorescence
Peripheral Blood Monocyte
45 min
2% Goat Serum in PBS for 15 min prior to FITC conjugated Primary antibody incubation
NA
Fluorescence Microscopy
Successfully able to image CD14 positive monocytes in mixed population of peripheral blood mononuclear cells isolated from fresh human blood using wide-field microscopy. Stained for nucleus using Draq5 to obtain per cell information Fixing and staining done in suspension. Plated after staining on Concanavalin A treated plate. Allowed to settle for 30 min and spin down the plate before imaging. I have tried using without block with any serum and found no non-specific staining compared to blocked cells. Using for microscopy, it makes it easy to monitor sub-cellular events in parallel by staining with second marker of interest.
Normally, this is used for flow cytometery and FACS sorting
Easy to use. Minimal non-specific staining.
None
Best antibody. Staining post-fix gives flexibility in terms of imaging 24-48 hrs after staining. It's a one step process.