Cd14 clone hcd14

Manufactured by BioLegend

Sourced in United States

CD14 (clone HCD14) is a cell surface receptor that serves as a co-receptor for the detection of bacterial lipopolysaccharide (LPS). It is primarily expressed on monocytes and macrophages.

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Lab products found in correlation

Cd4 clone rpa t4, by BioLegend (2 mentions) Cd235a clone hir2, by Thermo Fisher Scientific (1 mentions) Cd38 clone hit2, by BioLegend (1 mentions) Cd117 c kit, by Thermo Fisher Scientific (1 mentions) Cd34 clone 581, by BioLegend (1 mentions) Cd16 clone 3g8, by BioLegend (1 mentions) Facsdiva software, by BD (1 mentions) Facscelesta cell analyzer, by BD (1 mentions) Clone mφp9, by BD (1 mentions) Golgiplug, by BD (1 mentions) Live dead stain, by Thermo Fisher Scientific (1 mentions) Ficoll paque plus, by GE Healthcare (1 mentions) Clone mpc 11, by BioLegend (1 mentions) Clone mopc 21, by BioLegend (1 mentions) Il 8 clone e8n1, by BioLegend (1 mentions) Triton x 100, by Merck Group (1 mentions) Fix perm buffer, by Thermo Fisher Scientific (1 mentions) Paraformaldehyde (pfa), by Thermo Fisher Scientific (1 mentions) Sodium citrate, by BD (1 mentions) Hla dr clone l243, by BioLegend (1 mentions)

Spelling variants of this product

HCD14 (9 citations) CD14 (HCD14, (6 citations) Clone HCD14 (4 citations) PE-anti-CD14 (clone HCD14), (4 citations) CD14-PE/Cy7 (clone HCD14, (3 citations) FITC-labeled anti-CD14 (Clone HCD14) (2 citations) PerCP/Cy5.5-CD14 clone HCD14 (2 citations) Anti-CD14 (clone HCD14; (2 citations) CD14-FITC antibody (clone HCD14, (2 citations)

8 protocols using cd14 clone hcd14

Multiparameter Flow Cytometry of Hematopoiesis

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To observe erythroid differentiation in human tissues, staining was performed with a combination of CD117/c-Kit (clone YB5.B8; eBioscience), CD34 (clone 581; BioLegend), CD38 (clone HIT2; BioLegend), CD36 (clone 5-271; BioLegend), CD71 (clone OKT9; eBioscience), and CD235a (clone HIR2; eBioscience). To observe monocytic and granulocytic differentiation, antibodies against CD117/c-Kit, CD34, CD38, CD15 (clone HI98; eBioscience), CD14 (clone HCD14; BioLegend), and CD16 (clone 3G8; BioLegend) were used.

McKenney A.S., Lau A.N., Somasundara A.V., Spitzer B., Intlekofer A.M., Ahn J., Shank K., Rapaport F.T., Patel M.A., Papalexi E., Shih A.H., Chiu A., Freinkman E., Akbay E.A., Steadman M., Nagaraja R., Yen K., Teruya-Feldstein J., Wong K.K., Rampal R., Heiden M.G., Thompson C.B, & Levine R.L. (2018). JAK2/IDH-mutant–driven myeloproliferative neoplasm is sensitive to combined targeted inhibition. The Journal of Clinical Investigation, 128(2), 789-804.

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Multicolor Flow Cytometry of MSCs

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The surface antigen expression of MSCs was identified by flow cytometry using the following antibodies: CD14 (clone HCD14; BioLegend, San Diego, United States or clone MφP9; BD Biosciences, New Jersey, United States), CD34 (clone 8G12 also known as HPCA2), CD45 (clone HI30), CD73 (clone AD2), CD90 (clone 5E10), CD105 (clone 266), and HLA DRDPDQ (clone Tu39 also known as TÜ39) (all from BD Biosciences). The cells were stained as per manufacturer’s instructions (for staining details see Supplementary Table S1) and fluorescence intensities were measured using the FACSCelesta™ Cell Analyzer with BD FACSDiva™ software (BD Biosciences). Surface antigens were subdivided into identity markers (CD73, CD90, and CD105) and purity markers (CD14, CD34, CD45, and HLA DRDPDQ).

Jakl V., Ehmele M., Winkelmann M., Ehrenberg S., Eiseler T., Friemert B., Rojewski M.T, & Schrezenmeier H. (2023). A novel approach for large-scale manufacturing of small extracellular vesicles from bone marrow-derived mesenchymal stromal cells using a hollow fiber bioreactor. Frontiers in Bioengineering and Biotechnology, 11, 1107055.

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RRMS Patient PBMC Stimulation and Cytokine Analysis

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RRMS patients treated with natalizumab (n = 9) and healthy donors (n = 11) were recruited and PBMCs were isolated by density gradient centrifugation using Ficoll-Paque plus (GE Healthcare) and stored in liquid nitrogen until use. Upon thawing, cells were washed, counted, and resuspended in RPMI medium supplemented with 20% FCS, L-glutamine, and penicillin-streptomycin. Cells were plated at 2 × 10⁶ cells/mL and stimulated with 40 μg plate-bound TDB, 50 ng soluble LPS, or wells were treated with isopropanol prior to air drying as negative controls. Cells were cultured for 48 hours and treated with GolgiPlug (BD Biosciences) 5 hours prior to termination of the experiment. Cells were stained for MCL (clone 9B9, catalog 360204) and isotype (clone MPC-11, catalog 400314) expression prior to in vitro stimulations (BioLegend). Upon stimulation, cells were stained with LIVE/DEAD stain (Thermo Fisher Scientific, catalog L34976), CD14 (clone HCD14, catalog 325608), IL-8 (clone E8N1, catalog 511406), IL-6 (clone MQ2-13A5, catalog 501114), TNF (clone Mab11, catalog 502926), or isotype for cytokines (clone MOPC-21, catalog 400108) (BioLegend).

N’diaye M., Brauner S., Flytzani S., Kular L., Warnecke A., Adzemovic M.Z., Piket E., Min J.H., Edwards W., Mela F., Choi H.Y., Magg V., James T., Linden M., Reichardt H.M., Daws M.R., van Horssen J., Kockum I., Harris R.A., Olsson T., Guerreiro-Cacais A.O, & Jagodic M. (2020). C-type lectin receptors Mcl and Mincle control development of multiple sclerosis–like neuroinflammation. The Journal of Clinical Investigation, 130(2), 838-852.

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Peripheral Blood Cell Immunophenotyping in OA and RA

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We collected fresh peripheral blood from OA and RA patients with heparin sodium anticoagulation tubes and obtained peripheral blood mononuclear cells (PBMCs) using Ficoll-Paque gradient centrifugation. PBMCs were stained with fluorochrome-labelled anti-human monoclonal antibodies (Biolegend Inc., San Diego, CA) to CD45 (clone HI30, Cat No. 304014 / 304007 / 304005), CD14 (clone HCD14, Cat No. 325620), CD19 (clone SJ25C1, Cat No. 363006), CD3 (clone OKT3, Cat No. 317343), CD4 (clone RPA-T4, Cat No. 300538), CD8α (clone RPA-T8, Cat No. 301008), and CD16 (clone 3G8, Cat No. 302008) for 15 min at room temperature, followed by DAPI (Cat No. 422801) staining for 10 min. Using a flow cytometer, antibody-stained patient lymphocytes were sorted into monocytes (DAPI-CD45+CD3+CD19-CD14+), B cells (DAPI-CD45+CD3-CD19+), CD4+ T cells (DAPI-CD45+CD3+CD19-CD4+CD8-), and CD8+ T cells (DAPI-CD45+CD3+CD19-CD4+CD8+). At least 50,000 cells were enriched. Post-sort purities of each cell type were ensured for > 95% with flow cytometry. For monocyte subpopulation analysis, monocytes were classified as classical (DAPI-CD45+CD3+CD19-CD14++CD16-), intermediate (DAPI-CD45+CD3+CD19-CD14++CD16+), and non-classical (DAPI-CD45+CD3+CD19- CD14+CD16+).

Zong D., Huang B., Li Y., Lu Y., Xiang N., Guo C., Liu Q., Sha Q., Du P., Yu Q., Zhang W., Cai P., Sun Y., Tao J., Li X., Cai S, & Qu K. (2021). Chromatin accessibility landscapes of immune cells in rheumatoid arthritis nominate monocytes in disease pathogenesis. BMC Biology, 19, 79.

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Multiparametric Immunophenotyping of PBMCs

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Whole blood samples were collected in cell preparation tubes with sodium citrate (BD Biosciences). PBMCs were obtained by centrifugation and viably frozen until analysis. PBMCs were thawed, washed with flow buffer (5% BSA, 2 mM EDTA in PBS) and incubated with LIVE/DEAD Fixable Aqua Dead Cell Stain (Life Technologies), Fc receptor blocking agent (Miltenyi Biotec) and stained with surface antibodies (CD3 clone OKT3, CD4 clone RPA-T4, CD8 clone SK1, CD14 clone HCD14, CD19 clone HIB19, CD25 clone BC96, ICOS clone C398.4A, HLA-DR clone L243, PD-1 clone 29F.1A12 all from BioLegend) for 20 min at 4°C. For Foxp3 and Ki67 staining, cells were fixed and permeabilized using a Fix/Perm buffer (eBiosciences) according to the manufacturer’s instructions, then stained with anti-Foxp3 (clone 206D, BioLegend) or anti-Ki67 antibody (clone B56, BD Biosciences).
For global protein acetylation analysis, after surface staining, cells were fixed with 0.4% paraformaldehyde (Thermo Fisher Scientific), permeabilized in Triton X-100 (Sigma-Aldrich) and subsequently stained with anti-acetylated lysine antibody (clone 15G10, BioLegend).

Hellmann M.D., Jänne P.A., Opyrchal M., Hafez N., Raez L.E., Gabrilovich D., Wang F., Trepel J.B., Lee M.J., Yuno A., Lee S., Brouwer S., Sankoh S., Wang L., Tamang D., Schmidt E., Meyers M.L., Ramalingam S.S., Shum E, & Ordentlich P. (2020). Entinostat plus pembrolizumab in patients with metastatic NSCLC previously treated with anti-PD-(L)1 therapy. Clinical cancer research : an official journal of the American Association for Cancer Research, 27(4), 1019-1028.

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Characterizing Monocyte Activation by CAR T Cells

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CD14⁺ monocytes were isolated from PBMCs by positive selection. Isolated monocytes (10,000 cells) were plated in a 96-wells flat-bottom plate. 24 hours later, 10,000 CAR T cells and 10,000 tumor cells (MM1.S) were added in each well and were incubated for 24 hours. After incubation, the wells were washed with phosphate-buffered saline twice and detached from the wells with Trypsin/EDTA solution (Lonza) for 20 minutes. Monocytes were stained with CD3 (clone SK7, BD Biosciences), CD14 (clone HCD14, BioLegend), HLA-DR (clone G46-6, BD Biosciences), CD86 (clone HA5.2B7, Beckman Coulter), and CD80 (clone 16-10A1, BioLegend) (staining with 1 μg per mL of sample, incubation for 30 minutes at 4°C in PBS with 1% BSA, 10% FBS and 0.1% NaN3 sodium azide). Flow cytometry was performed on BD LSR Fortessa.

Katsarou A., Sjöstrand M., Naik J., Mansilla-Soto J., Kefala D., Kladis G., Nianias A., Ruiter R., Poels R., Sarkar I., Patankar Y.R., Merino E., Reijmers R.M., Frerichs K.A., Yuan H., de Bruijn J., Stroopinsky D., Avigan D., van de Donk N.W., Zweegman S., Mutis T., Sadelain M., Groen R.W, & Themeli M. (2021). Combining a CAR and a chimeric costimulatory receptor enhances T cell sensitivity to low antigen density and promotes persistence. Science translational medicine, 13(623), eabh1962.

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Multicolor Flow Cytometry Staining

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Dead cells were excluded by the fixable viability dye eFluor 780 (eBioscience) and FcR-dependent staining was blocked with human FcR blocking reagent (Miltenyi). Cells were stained with fluorophore-conjugated antibodies against CD45 (clone HI30, BioLegend), CD14 (clone HCD14, BioLegend), HLA-DR (clone L242, BioLegend), CD1a (clone HI149, BioLegend), human Langerin (clone MB22-9F5, Miltenyi) for either 30 min or 15 min on ice. Experiments were conducted with an Attune NxT flow cytometer (ThermoScientific) or FACS Canto II (BD Biosciences) and analysed in FlowJo software.

Rentzsch M., Wawrzinek R., Zelle-Rieser C., Strandt H., Bellmann L., Fuchsberger F.F., Schulze J., Busmann J., Rademacher J., Sigl S., Del Frari B., Stoitzner P, & Rademacher C. (2021). Specific Protein Antigen Delivery to Human Langerhans Cells in Intact Skin. Frontiers in Immunology, 12, 732298.

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Multicolor Flow Cytometry Antibody Panel

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The following antimouse antibodies were purchased from BioLegend: CD45 [clone 30-F11; PerCP5.5 or phycoerythrin (PE)–Cyanine7], T cell receptor (TCR) β chain (clone H57-597; PerCP5.5), TCR γ/δ (clone GL3; PE-Cyanine7), CD11b (clone M1/70; Pacific Blue), F4/80 [clone BM8; allophycocyanin (APC) or PerCP5.5], CD127 (clone A7R34; Brilliant Violet 605), CD90.2 (clone 30-H12; Alexa Fluor 488), and IL-17A [clone TC11-18H10.1; fluorescein isothiocyanate (FITC) or APC]. PE streptavidin and antibodies for CD16/CD32 (clone 2.4G2; purified), CD3ε (clone 145-2C11; PerCP5.5), Gr-1 (clone RB6-8C5; APC), and RORγt (clone Q31-378; Alexa Fluor 647 or BV421) were purchased from BD Pharmingen. Antibodies for TNF-α (clone TN3-19; PE-Cyanine7) and mouse anti-iNOS (clone 6/iNOS/NOS Type II; FITC) antibodies were purchased from BD Transduction Laboratories.
The antihuman lineage cocktail, including CD3, CD14, CD16, CD19, CD20, and CD56 (clones UCHT1, HCD14, 3G8, HIB19, 2H7, and HCD56; APC), CD244 (clone C1.7; PE), CD11b (clone CBRM1/5; PerCP-Cy5.5), HLA-DR (clone L243; Brilliant Violet 785), CD14 (clone HCD14; Brilliant Violet 421), CD33 (clone P67.6; APC/Cy7), CD11c (clone 3.9; APC), CD80 (clone 2D10; PE-Cy7), and CD86 (clone IT2.2; Brilliant Violet 711), were purchased from BioLegend. Antibodies for human NOS2 (clone C-11; Alexa Fluor 488) were purchased from Santa Cruz Biotechnology.

Zhong J., Scholz T., Yau A.C., Guerard S., Hüffmeier U., Burkhardt H, & Holmdahl R. (2018). Mannan-induced Nos2 in macrophages enhances IL-17–driven psoriatic arthritis by innate lymphocytes. Science Advances, 4(5), eaas9864.

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