Anti human cd14 clone 63d3

Manufactured by BioLegend

Anti-human CD14 (clone 63D3) is a monoclonal antibody that binds to the CD14 protein, which is a receptor for lipopolysaccharide (LPS) and plays a role in the innate immune response.

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Lab products found in correlation

Lsrfortessa, by BD (1 mentions) Anti human cd16 clone 3g8, by BioLegend (1 mentions) Cytoflex flow cytometer, by Beckman Coulter (1 mentions) Anti human cd3 clone okt3, by BioLegend (1 mentions) Attune nxt, by Thermo Fisher Scientific (1 mentions) Accutase, by STEMCELL (1 mentions)

Close spelling variants of this product

Anti-CD14 (clone 63D3; (3 citations) CD14 (63D3; (3 citations) Clone 63D3 (2 citations)

3 protocols using anti human cd14 clone 63d3

Flow Cytometry Analysis of Hematopoietic Markers

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For analysis of surface markers, cells were stained in PBS containing 2% (w/v) BSA, with the following antibodies (from Biolegend): anti-human CD34 (clone 581, Cat# 343504, 1:200), anti-human CD33 (clone P67.6, Cat# 366608, 1:200), anti-human CD117 (clone 104D2, Cat# 313205, 1:200), anti-human CD16 (clone 3G8, Cat# 302046, 1:100), anti-human CD14 (clone 63D3, Cat# 367117, 1:200). Monocyte and macrophage were indicated by anti-human CD14⁺. Flow cytometry data were acquired on a LSRII or LSR Fortessa (BD Biosciences) and analyzed using FlowJo software (Tree Star).

Wheeler M.M., Stilp A.M., Rao S., Halldórsson B.V., Beyter D., Wen J., Mihkaylova A.V., McHugh C.P., Lane J., Jiang M.Z., Raffield L.M., Jun G., Sedlazeck F.J., Metcalf G., Yao Y., Bis J.B., Chami N., de Vries P.S., Desai P., Floyd J.S., Gao Y., Kammers K., Kim W., Moon J.Y., Ratan A., Yanek L.R., Almasy L., Becker L.C., Blangero J., Cho M.H., Curran J.E., Fornage M., Kaplan R.C., Lewis J.P., Loos R.J., Mitchell B.D., Morrison A.C., Preuss M., Psaty B.M., Rich S.S., Rotter J.I., Tang H., Tracy R.P., Boerwinkle E., Abecasis G.R., Blackwell T.W., Smith A.V., Johnson A.D., Mathias R.A., Nickerson D.A., Conomos M.P., Li Y., Þorsteinsdóttir U., Magnússon M.K., Stefansson K., Pankratz N.D., Bauer D.E., Auer P.L, & Reiner A.P. (2022). Whole genome sequencing identifies structural variants contributing to hematologic traits in the NHLBI TOPMed program. Nature Communications, 13, 7592.

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NECA Modulation of Human PBMC Activation

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Purified human PBMCs from three healthy donors were co-cultured with various concentrations of NECA and were stimulated with anti-human CD3 and CD28 antibody (at 1 μg/mL). Cells were kept in culture for 48 hours days. Golgi block was added 4 hours prior to intracellular flow cytometry analysis. Antibodies used in analysis include: anti-human CD8a (Clone RPA-T8, BioLegend Cat No. 301048), anti-human CD3 (Clone OKT3, BioLegend Cat No. 317322), anti-human CD4 (clone OKT4, BioLegend Cat No. 317436), anti-human CD56 (clone 5.1H11, BioLegend Cat No. 362546), anti-human CD205 (clone HD30, BioLegend Cat No. 342210) anti-human CD14 (clone 63D3, BioLegend Cat No. 367118), anti-human CD19 (clone SJ25C1, BioLegend Cat No. 363034), anti-human CXCL5 (clone J111B7, BioLegend Cat No. 524104), anti-human MCP-1 (clone 2H5, BioLegend Cat No. 505904), anti-human IL-8 (clone E8N1, BioLegend Cat No. 511408), anti-human CXCL1 (clone 20326, R&D Systems Cat No. IC275P). Data was acquired on a CytoFLEX flow cytometer (Beckman Coulter) and analyzed in FlowJo software v10.

Fong L., Hotson A., Powderly J.D., Sznol M., Heist R.S., Choueiri T.K., George S., Hughes B.G., Hellmann M.D., Shepard D.R., Rini B.I., Kummar S., Weise A.M., Riese M.J., Markman B., Emens L.A., Mahadevan D., Luke J.J., Laport G., Brody J.D., Hernandez-Aya L., Bonomi P., Goldman J.W., Berim L., Renouf D.J., Goodwin R.A., Munneke B., Ho P.Y., Hsieh J., McCaffery I., Kwei L., Willingham S.B, & Miller R.A. (2019). Adenosine A2A Receptor Blockade as an Immunotherapy for Treatment-Refractory Renal Cell Cancer. Cancer discovery, 10(1), 40-53.

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Phagocytosis Assay for Macrophages

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Monocyte-derived macrophages were obtained by culturing purified monocytes in RPMI 10% FBS for 7 days. Non-adherent cells are removed. Phagocytosis assay was performed by incubating DAPI-treated trophozoites at a ratio of 5 iRBCs to 1 macrophage for 90 mins. Thereafter, the co-culture was washed in PBS to remove excess iRBCs. Macrophages were detached using Accutase (StemCell Technologies), stained with anti-human CD14 (Clone 63D3; Biolegend, 1:100 dilution), and quantified using Attune NxT (Life Technologies) and analyzed using Flowjo v10. Percentage phagocytosis is calculated as the percentage of DAPI-positive macrophages.

Chew M., Ye W., Omelianczyk R.I., Pasaje C.F., Hoo R., Chen Q., Niles J.C., Chen J, & Preiser P. (2022). Selective expression of variant surface antigens enables Plasmodium falciparum to evade immune clearance in vivo. Nature Communications, 13, 4067.

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