BioLegend
PerCP/Cy5.5 anti-human CD68 Antibody
333814
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One of the general aims of our lab is to help characterize and understand the role that macrophage and monocyte subsets have in disease. Several different subsets of macrophages and monocytes have been described in the literature over the past few years, with flow cytometry being the main tool used to characterize the phenotype of these cells. One important marker for macrophage/monocytes is CD68. Quality antibodies are required for successful characterization of rare cell subsets, especially in the context of multi-parameter flow cytometry. The purpose of this experiment was to identify an anti-CD68 antibody for use with human cells. This particular product was selected for testing because it is (a) directly labelled with a fluorophore, (b) already titrated for use in flow cytometry, and (c) BioLegend has an excellent reputation for their antibodies.
Flow Cytometry
human peripheral blood mononuclear cells isolated from whole blood by Ficoll gradient separation
Fix cells on ice for 20 minutes with 4% paraformaldehyde diluted in PBS. Wash twice with FACS buffer (PBS+2% FCS, 1mM EDTA, 0.1% sodium azide). Permeabilize with 0.5% saponin diluted in FACS buffer for 30 minutes on ice. Wash cells twice with FACS buffer
N/A
Cells were simultaneously incubated with a FITC anti-human CD3 antibody and PerCP-Cy5.5-anti-human CD68 (333814, clone Y1/82A) for 1 hour on ice at 1:40 dilution in a total volume of 100ul FACS buffer.
BD FACSCalibur
As expected, CD68 staining was restricted to CD3- cells. CD68 staining was bright, with approximately 40% of CD3- cells expressing CD68.
For analysis, quadrant gates were set so that >99% of events were negative when stained with a matched isotype control.
Bright staining. Antibody comes pre-diluted ready to use. Directly-labelled with a fluorophore.
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If you are looking for an anti-CD68 antibody for human samples, you can't get much better than this product!