Anti human cd14 pacificblue clone m5e2

Manufactured by BD

Sourced in Germany

Anti-human CD14-PacificBlue (clone M5E2) is a fluorochrome-conjugated monoclonal antibody that recognizes the CD14 antigen expressed on the surface of human monocytes, macrophages, and some dendritic cells. It can be used for the identification and enumeration of these cell populations in flow cytometric applications.

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Lab products found in correlation

Anti human hla dr apccy7, by BioLegend (1 mentions) Lsrfortessa, by BD (1 mentions)

3 protocols using anti human cd14 pacificblue clone m5e2

Multiparametric Flow Cytometry Immunophenotyping

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For flowcytometric analysis, harvested cells were stained for 20 min at 4 °C using the following antibodies: anti-human CD3-Allophycocyanin (APC) (clone UCHT1, BD Pharmingen, San Jose, CA, USA), anti-human CD4- Phycoerythrin (PE) (clone RPA-T4, BD Pharmingen), anti-human CD8-PacificBlue (clone RPA-T8, BD Pharmingen), anti-human CD14-PacificBlue (clone M5E2, BD Pharmingen), anti-human CD40-PacificBlue (clone 5C3, BioLegend, Fell, Germany), anti-human CD69-APC-Cyanine (Cy)7 (clone FN50, BioLegend), anti-human CD80-APC (clone MEM-233, EuroBioSciences, Friesoythe, Germany), anti-human CD83- Fluorescein (FITC) (clone HB15e, BD Pharmingen), anti-human CD86-PE (clone IT2.2, BioLegend), anti-human HLA-ABC-PacificBlue (clone W6/32, BioLegend), and anti-human HLA-DR-APCCy7 (clone L243, BioLegend). Flowcytometric analyses were performed using a LSRFortessa flow cytometer. Data were analyzed using the BD FACSDiva software version 8.0.3 (BD Biosciences, Heidelberg, Germany) or FlowJo software version 10.7.1 (Tree Star, Ashland, Wilmington, DE, USA).

Krauter S., Büscher N., Bräuchle E., Ortega Iannazzo S., Penner I., Krämer N., Gogesch P., Thomas S., Kreutz M., Dejung M., Freiwald A., Butter F., Waibler Z, & Plachter B. (2022). An Attenuated Strain of Human Cytomegalovirus for the Establishment of a Subviral Particle Vaccine. Vaccines, 10(8), 1326.

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Clever-1 Expression on PBMCs

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Peripheral blood mononuclear cells were plated at 0.2×10⁶ cells/well in round-bottom 96-well plates. All wells were stained with anti-human CD14-Pacific Blue (clone M5E2, BD Pharmingen) together with 10 ng/μL in-house conjugated (AF647) anti-Clever-1 antibody bexmarilimab (competitor). In-house conjugated Irrelevant isotype control human IgG4 (S241/L248E) was used for signal normalization. Flow cytometry was run on the LSRFortessa (BD) and analyzed with FlowJo software v10.7.1 (BD).

Rannikko J.H., Verlingue L., de Miguel M., Pasanen A., Robbrecht D., Skytta T., Iivanainen S., Shetty S., Ma Y.T., Graham D.M., Arora S.P., Jaakkola P., Yap C., Xiang Y., Mandelin J., Karvonen M.K., Jalkanen J., Karaman S., Koivunen J.P., Minchom A., Hollmén M, & Bono P. (2023). Bexmarilimab-induced macrophage activation leads to treatment benefit in solid tumors: The phase I/II first-in-human MATINS trial. Cell Reports Medicine, 4(12), 101307.

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Comprehensive Immune Cell Profiling

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PBMCs were plated at 0.2×10⁶ cells/well in round-bottom 96-well plates. All wells were stained with anti–human CD14-Pacific Blue (clone M5E2; BD Pharmingen) together with 10 ng/μL in-house conjugated (AF647) anti–Clever-1 antibodies 9-11 or FP-1305. Irrelevant isotype control antibodies rat IgG2a or human in-house conjugated IgG4 (S241/L248E) were used for signal normalization, respectively. Blood neutrophils were identified by anti–CD11b-AF488 (clone ICRF44; BioLegend) and anti–CD15-APC (clone HI98; BD) after red blood cell (RBC) lysis with PharmLyse. Lymphocyte FACS was performed by Covance according to their standard protocols.

Virtakoivu R., Rannikko J.H., Viitala M., Vaura F., Takeda A., Lönnberg T., Koivunen J., Jaakkola P., Pasanen A., Shetty S., de Jonge M.J., Robbrecht D., Ma Y.T., Skyttä T., Minchom A., Jalkanen S., Karvonen M.K., Mandelin J., Bono P, & Hollmén M. (2021). Systemic Blockade of Clever-1 Elicits Lymphocyte Activation Alongside Checkpoint Molecule Downregulation in Patients with Solid Tumors: Results from a Phase I/II Clinical Trial. Clinical Cancer Research, 27(15), 4205-4220.

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