Antibodies directed against cd14

Manufactured by Miltenyi Biotec

Antibodies directed against CD14 are laboratory reagents used for the detection and analysis of the CD14 protein. CD14 is a cell surface receptor that plays a role in the innate immune response. These antibodies can be used to identify and quantify cells expressing CD14, such as monocytes and macrophages, through techniques like flow cytometry.

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Lab products found in correlation

Rpmi 1640, by Thermo Fisher Scientific (3 mentions) Macrophage colony stimulating factor m csf, by Thermo Fisher Scientific (2 mentions) Alexa 555, by Cell Signaling Technology (1 mentions) Alexa fluor 488 phalloidin, by Thermo Fisher Scientific (1 mentions) Mouse anti vinculin clone hvin 1, by Merck Group (1 mentions) Glass coverslip, by Marienfeld (1 mentions) Dt 1bon1000 96, by Immunodiagnostic Systems (1 mentions) Rankl, by Miltenyi Biotec (1 mentions) M csf, by Thermo Fisher Scientific (1 mentions) Accutase solution, by Merck Group (1 mentions) Magnetic microbead, by Miltenyi Biotec (1 mentions) Trypsin edta, by Thermo Fisher Scientific (1 mentions)

3 protocols using antibodies directed against cd14

Isolation and Differentiation of Human Monocytes

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Human monocytes were isolated from blood of healthy donors as described previously (Van Goethem et al., 2010 (link)). Cells were re-suspended in cold PBS supplemented with 2 mM EDTA, 0.5% heat-inactivated fetal calf serum (FCS) at pH 7.4 and magnetically sorted with magnetic microbeads coated with antibodies directed against CD14 (Miltenyi Biotec). Monocytes were then seeded on glass coverslips at 1.5x10⁶ cells/well in six-well plates in RPMI 1640 (Invitrogen) without FCS. After 2 h at 37°C in humidified 5% CO₂ atmosphere, the medium was replaced by RPMI containing 10% FCS and 20 ng/mL of Macrophage Colony-Stimulating Factor (M-CSF) (Peprotech). For experiments, cells were used after seven days of differentiation.
Macrophages plated on glass coverslips (0.17 mm ±0.005mm, Marienfeld) were unroofed and fixed as previously described (Bouissou et al., 2017 (link)) and labelled with Alexa Fluor 488-phalloidin (Molecular Probes, 1/500) and Alexa Fluor 647-coupled phalloidin (Molecular Probes, A22287, 1/100) for RIM and dSTORM, respectively. Vinculin was stained using mouse anti-vinculin (clone hvin-1, Sigma-Aldrich, 1/500) and a secondary F(ab')₂ coupled to Alexa 555 (Cell Signaling technology).

Mangeat T., Labouesse S., Allain M., Negash A., Martin E., Guénolé A., Poincloux R., Estibal C., Bouissou A., Cantaloube S., Vega E., Li T., Rouvière C., Allart S., Keller D., Debarnot V., Wang X.B., Michaux G., Pinot M., Le Borgne R., Tournier S., Suzanne M., Idier J, & Sentenac A. (2021). Super-resolved live-cell imaging using random illumination microscopy. Cell Reports Methods, 1(1), 100009.

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Isolation and Differentiation of Human Monocytes

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Human monocytes were isolated from blood of healthy donors as previously described (Van Goethem et al., 2010 (link)). Cells were re-suspended in cold PBS supplemented with 2 mM EDTA, 0.5% heat-inactivated fetal calf serum (FCS) at pH 7.4 and magnetically sorted with using magnetic microbeads coated with antibodies directed against CD14 (Miltenyi Biotec). Monocytes were then seeded on plastic at 2 × 10⁶ cells/well in six-well plates in RPMI 1640 (Invitrogen) without FCS. After 2 h at 37 °C in humidified 5% CO₂ atmosphere, the medium was replaced by RPMI containing 10% FCS, 100 ng/mL of macrophage colony-stimulating factor (M-CSF, Peprotech) and 60 ng/mL of human receptor activator of NF-κB-ligand (RANK-L, Miltenyi Biotec). The medium was then changed every third day by RPMI containing 10% FCS, 100 ng/mL of RANK-L and 25 ng/mL of M-CSF. For experiments, cells were harvested at day 10 using Accutase solution (Sigma-Aldrich) and centrifugation (1100 rpm, 10 min), and were then plated either on clean 1.5 H precision glass coverslips (Marienfeld 0117640) or on bovine bone slices (Immuno Diagnostic Systems DT-1BON1000-96). Cells were left to adhere in cytokine-supplemented medium for 3 days before fixation.

Portes M., Mangeat T., Escallier N., Dufrancais O., Raynaud-Messina B., Thibault C., Maridonneau-Parini I., Vérollet C, & Poincloux R. (2022). Nanoscale architecture and coordination of actin cores within the sealing zone of human osteoclasts. eLife, 11, e75610.

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Isolation and Culture of Human Monocytes

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Human monocytes were isolated from the blood of healthy donors as described previously 37 (link) .
Cells were resuspended in cold phosphate buffered saline (PBS) supplemented with 2 mM EDTA, 0.5% heat-inactivated Fetal Calf Serum (FCS) at pH 7.4 and magnetically sorted with magnetic microbeads coated with antibodies directed against CD14 (Miltenyi Biotec). Monocytes were then seeded on glass coverslips at 1.5 × 10 6 cells/well in six-well plates in RPMI 1640 (Invitrogen) without FCS. After 2h at 37°C in a humidified 5% CO2 atmosphere, the medium was replaced by RPMI containing 10% FCS and 20 ng/mL of Macrophage Colony-Stimulating Factor (M-CSF) (Peprotech). For experiments, cells were harvested at day 7 using trypsin-EDTA (Fisher Scientific) and centrifugation (320g, 10 min).

Jasnin M., Hervy J., Balor S., Proag A., Proag A., Voituriez R., Maridonneau‐Parini I., Baumeister W., Dmitrieff S., & Poincloux R. (2021). Elasticity of dense actin networks produces nanonewton protrusive forces.

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