CD14 FITC Antibody Using Human PBMCs

October 22, 2015

Anaesthesiology
LMU Munich
Researcher

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Company:

BioLegend

Product Name:

Anti human CD14 FITC

Catalog Number:

325603

Info: View Product View Product Specs

Browse Similar Products: CD14 Antibodies

We were searching for a reliable antibody for identifying CD14 positive cells from PBMCs, such as monocytes etc. FITC-conjugated antibodies proved to be reliable. In the literature, there is a body of evidence that Cy-colors tend to bind unspecifically to monocytes and macrophages via Fc Receptor binding, therefore we chose FITC.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Human PBMCs

Primary Incubation

10 mins room temperature (Fc Receptor blocking, 5µl for a 100µl cell suspension containing 2 milllion cells)

Blocking Agent

TruStain FcX blocking solution (BioLegend)

Secondary Incubation

2µl of antibody on 2 millions cells, 30 mins on ice in the dark (antibody), wash three times afterwards with 2ml of FACS buffer

Tertiary Incubation

None

Detection

Flow Cytometry

Results Summary

This proved to be a relaible antibody for gating on CD14 positive cells from a suspension of PBMCs (CD14 positive cells such as monocytes etc.) Together with CD16 gating, a robust panel of monocyte phenotyping may be created.

Additional Notes

Be sure to always block with a blockig reagent due to unselective binding of the antibodies to monocytes due to Fc Receptors. Due to this matter, always compare this antibody to the corresponding Isotype.

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Summary

The Good

Quick incubation time, fluorescently labeled, not much antibody solution needed, affordable

The Bad

Washing time needed (at least for PBMCs), but that's all

The Bottom Line

Quick, reliable staining for phenotyping of CD14 positive populations in PBMCs, macrophages, etc. Easy to use, affordable

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