Pe mouse anti human cd14

Manufactured by BD

Sourced in United States

PE mouse anti-human CD14 is a laboratory reagent that detects the CD14 antigen expressed on the surface of human cells. CD14 is a glycosylphosphatidylinositol-anchored protein that serves as a receptor for lipopolysaccharide. This product can be used in flow cytometry and other immunoassays to identify and quantify CD14-positive cells.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

CD14-PE (78 citations) Anti-CD14-PE (40 citations) Mouse anti (2 citations)

11 protocols using pe mouse anti human cd14

Isolation and Enrichment of Human CD14+ Monocytes

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human PBMCs were isolated using lymphocyte separation media (MP Biomedicals, Solon, OH) as directed by the manufacturer. In brief, blood was collected from normal healthy donors by venipuncture in 10 mL EDTA Vacutainer tubes (Becton Dickinson, Franklin Lakes, NJ). After centrifugation the buffy coat was removed, resuspended in RPMI 1640 media supplemented with 10% FBS (Atlas Biologicals, Fort Collins, CO), 2 mM L-glutamine and antibiotics and layered over lymphocyte separation media. Cells were centrifuged for 30 min at 1000× g at 4°C, the buffy coat was removed, washed and cells were counted using a hemocytometer.
Human CD14⁺ monocytes were enriched by positive selection from PBMCs using CD14⁺ MACS technology (Miltenyi Biotec, Auburn, CA) as directed by manufacturer. CD14⁺ monocyte isolation was confirmed by flow cytometry using mouse anti-human CD14⁺ PE (BD Pharmingen, San Jose, CA) and CD14⁺ cells were routinely enriched to a purity of 94–98%.

Weinkopff T., Mackenzie C., Eversole R, & Lammie P.J. (2014). Filarial Excretory-Secretory Products Induce Human Monocytes to Produce Lymphangiogenic Mediators. PLoS Neglected Tropical Diseases, 8(7), e2893.

+ Open protocol

+ Expand

Flow Cytometry Analysis of Monocyte Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

Blood samples (100 μl) were incubated for 30 minutes at room temperature in darkness with mouse anti-human CD14 PE (Clone TÜK4, BD Bioscience), mouse anti-human HLA-DR PerCP-Cy5.5 (Clone HL-38, Novus Biologicals), or polyclonal rabbit anti-human TLR2 Alexa Fluor 700 (Bioss Antibodies, 5 μl). Control samples were incubated with suggested isotype controls for the settings. After incubation, 3 ml of FACS lysing solution (BD Pharmingen^TM) were added and incubation for 10 minutes at room temperature in darkness followed. Subsequently, the samples were centrifuged at 800G for 5 minutes at room temperature. The pellet was resuspended in 500 μl of FACS buffer. The samples were measured by flow cytometry with BD FACS Canto^TM using FACS DIVA^TM software (BD Biosciences). The monocytes were discriminated by gating CD14⁺ cells. 300,000 monocytes were measured at least from each sample. Unstimulated samples were measured as control.

Schimunek L., Serve R., Teuben M.P., Störmann P., Auner B., Woschek M., Pfeifer R., Horst K., Simon T.P., Kalbitz M., Sturm R., Pape H.C., Hildebrand F., Marzi I, & Relja B. (2017). Early decreased TLR2 expression on monocytes is associated with their reduced phagocytic activity and impaired maturation in a porcine polytrauma model. PLoS ONE, 12(11), e0187404.

+ Open protocol

+ Expand

Liver Cell Isolation and Immune Cell Purification

Cited 3 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

The resected livers were cut into pieces and resuspended with a digestion enzyme mixture (2 mg/ml collagenase I, Invitrogen; 1 mg/mldispaseII, Sigma). The mixture of tissue/enzyme was put in a 37 °C shaker for 30 min and then poured through 70 μm cell strainers (BD Biosciences). After removing the hepatocytes using a Ficoll gradient (Percoll^™ PLUS/Percoll, GE) to obtain single-cell suspensions, flow cytometry analysis and fluorescence-activated cell sorting was fulfilled using fluorescently labeled antibodies (PE mouse anti-human CD14, PerCP-Cy^™5.5 mouse anti-human HLA-DR; PerCP-CyTM5.5 rabbit anti-mouse CD11b and APC rabbit anti-mouse Ly-6G and Ly-6C, BD). The PBMCs were separated from the peripheral blood of the patients by density gradient centrifugation (Percoll^™ PLUS/Percoll, GE)^{25 (link),29 (link),54 (link)}.

Lin Q., Ren L., Jian M., Xu P., Li J., Zheng P., Feng Q., Yang L., Ji M., Wei Y, & Xu J. (2019). The mechanism of the premetastatic niche facilitating colorectal cancer liver metastasis generated from myeloid-derived suppressor cells induced by the S1PR1–STAT3 signaling pathway. Cell Death & Disease, 10(10), 693.

+ Open protocol

+ Expand

Nanoparticle Synthesis and Characterization

Check if the same lab product or an alternative is used in the 5 most similar protocols

PLGA (75:25 L:G; ester-terminated, inherent viscosity range: 0.55–0.75 dL/g in CHCl₃) was purchased from Lactel. All lipids for the nanoparticle synthesize were purchased from Avanti Polar Lipids, including 1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOTAP), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[maleimide(polyethylene glycol)-2000] (DSPE-PEG-MAL). Cholesteryl butyrate was purchased from Santa Cruz Biotechnology. Rhesus recombinant anti-CD4 antibody and rhesus recombinant IgG1 isotype control antibody were purchased from NIH Nonhuman Primate Reagent Resource. FITC mouse anti-human CD8, PE mouse anti-human CD14, PerCP mouse anti-human CD3 antibodies, FITC mouse anti-human CD4, and FITC mouse IgG1, PE mouse IgG2a, PerCP mouse IgG1 κ control antibodies were purchased from BD Biosciences. RPMI 1640 containing 2 mMl-glutamine and 25 mM HEPES, DPBS, heat-inactivated fetal bovine serum (FBS), Penicillin–Streptomycin (10,000 U/mL), 1,1′ -Dioctadecyl-3,3,3′, 3′ -Tetramethylin-dodicarbocyanine, 4-Chlorobenzenesulfonate Salt (DiD), LIVE/DEAD^® Fixable Violet Dead Cell Stain Kit, Dylight 633 NHS Ester were purchased from ThermoFisher. All other chemicals were purchased from Sigma-Aldrich and Fisher Scientific unless otherwise specified.

Cao S., Jiang Y., Levy C.N., Hughes S.M., Zhang H., Hladik F, & Woodrow K.A. (2018). Optimization and comparison of CD4-targeting lipid–polymer hybrid nanoparticles using different binding ligands. Journal of biomedical materials research. Part A, 106(5), 1177-1188.

+ Open protocol

+ Expand

Multicolor Flow Cytometry Analysis of Monocytes

Check if the same lab product or an alternative is used in the 5 most similar protocols

U937 cells and PBMCs were analyzed on FACSCalibur flow cytometer (BD Biosciences). Fc receptors were preblocked with 10% FCS-containing DPBS for 30 min, 4°C. Cells were labeled with PE Mouse Anti-Human CD11b/Mac-1 (BD Pharmingen, #555388, 10 µg/ml), PE Mouse Anti-Human CD14 (BD Pharmingen, # 555398, 10 µg/ml) or PE Mouse IgG1 κ Isotype control (BD Pharmingen, #555749, 10 µg/ml) for 45 min, 4°C in the dark, thoroughly washed and analyzed.

Frank-Bertoncelj M., Pisetsky D.S., Kolling C., Michel B.A., Gay R.E., Jüngel A, & Gay S. (2018). TLR3 Ligand Poly(I:C) Exerts Distinct Actions in Synovial Fibroblasts When Delivered by Extracellular Vesicles. Frontiers in Immunology, 9, 28.

+ Open protocol

+ Expand

Flow Cytometric Analysis of PBMCs

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

PBMC were analyzed using a 3-laser BD LSRII flow cytometer and FlowJo software (Treestar) as described (9 (link)). Primary antibodies were anti-Apc1 (Novus; NBP1-77375), anti-PGK1 (Abcam; ab67335), anti-γH2AX (Cell Signaling; #9718), and anti-phospho-NF-κB p65 (Ser536, P-p65) (Cell Signaling; #3033). Secondary antibodies were FITC goat anti-mouse Ig and FITC goat anti-rabbit Ig (BD Biosciences). Cell surface markers (BD Biosciences) were Pacific Blue mouse anti-human CD4, Alexa Fluor 700 mouse anti-human CD8, APC mouse anti-human CD19, and PE mouse anti-human CD14. Mouse anti-human HLA-DR was from BD Pharmingen (G46-6). Cyclosporin A (30024), Bay 11-7085 (B5681), and BI-78D3 (B3063) were from Sigma-Aldrich.

Spurlock CF I.I.I., Tossberg J.T., Olsen N.J, & Aune T.M. (2015). Chronic NF-κB activation in CD4+ T cells in rheumatoid arthritis is genetically determined by HLA risk alleles. Journal of immunology (Baltimore, Md. : 1950), 195(3), 791-795.

+ Open protocol

+ Expand

Isolation and Analysis of Peripheral Blood Mononuclear Cells and Neutrophils

Check if the same lab product or an alternative is used in the 5 most similar protocols

Peripheral blood collected in the ethylene diamine tetraacetic acid (EDTA) tubes was 1:1 diluted with phosphate buffer saline (PBS) buffer, laid on the Ficoll density gradient (DAKEWE, China), and then centrifuged at 1800 rpm, 20 min, 24 °C (Figure S1). The PBMC were harvested at the interface layer, washed with PBS, and counted by cellmeter Auto T4 (Nexcelom Bioscience, USA). The percentage of low-density granulocytes (LDGs) were determined by PE Mouse Anti-Human CD14 (BD Biosciences, USA) and Percp-Cy5.5 Mouse Anti-Human CD15 (BD Biosciences, USA) staining (CD14^-/loCD15⁺).
Neutrophils were isolated from a red cell layer, followed by lysing buffer (BD Biosciences, USA) to remove red cells, washed with PBS, and counted. The purity of neutrophils was determined by CD16 staining (APC-conjugated anti-human CD16, BD Biosciences, USA) using flow cytometry (BD FASC arial II cytometer).

Peng Y., Wu X., Zhang S., Deng C., Zhao L., Wang M., Wu Q., Yang H., Zhou J., Peng L., Luo X., Chen Y., Wang A., Xiao Q., Zhang W., Zhao Y., Zeng X, & Fei Y. (2022). The potential roles of type I interferon activated neutrophils and neutrophil extracellular traps (NETs) in the pathogenesis of primary Sjögren’s syndrome. Arthritis Research & Therapy, 24, 170.

+ Open protocol

+ Expand

Flow Cytometric Immunophenotyping of CD19+ B Cells and CD14+ Monocytes

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were harvested, washed with PBS, and blocked with Human FC Receptor Binding Inhibitor (ThermoFisher, 16–9161). Cells were incubated with antibodies, APC-cy7 Mouse Anti-Human CD19 (BD Pharmingen, 557791), PE Mouse Anti-Human CD14 (BD Pharmingen, 555398). After washing with PBS, cells were resuspended in 7AAD (ThermoFisher, 00–6993) diluted with PBS. The analysis was performed on BD Accuri C6 flow cytometer (BD Biosciences) according to standard protocols with following changes in filters. We used filter 675/25 at F3 and 670/LP at F4.

Choi J., Lysakovskaia K., Stik G., Demel C., Söding J., Tian T.V., Graf T, & Cramer P. (2021). Evidence for additive and synergistic action of mammalian enhancers during cell fate determination. eLife, 10, e65381.

+ Open protocol

+ Expand

Characterization of Nanoformulated HIV Therapy

Check if the same lab product or an alternative is used in the 5 most similar protocols

DTG was a generous gift from ViiV Healthcare (Research Triangle Park, NC). Pyridine, dimethylformamide (DMF), N,N-diisopropylethylamine (DIEA), myristoyl chloride, poloxamer 407 (P407), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer, ciprofloxacin, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), paraformaldehyde (PFA), and 3,3’-diaminobenzidine (DAB) were purchased from Sigma-Aldrich (St. Louis, MO). Diethyl ether, cell culture grade water (endotoxin-free), gentamicin, acetonitrile (ACN), methanol, KH₂PO₄, bovine serum albumin (BSA), Triton X-100, LC-MS-grade water, and TRIzol reagent were purchased from Fisher Scientific (Hampton, NH). FITC mouse anti-human CD45, Alexa Fluor 700 mouse anti-human CD3, APC mouse anti-human CD4, BV421 mouse anti-human CD8, PE mouse anti-human CD14, and PE-Cy5 mouse anti-human CD19 were purchased from BD Biosciences (San Jose, CA). Monoclonal mouse anti-human HIV-1p24 (clone Kal-1), monoclonal mouse anti-human leukocyte antigen (HLA-DP/DQ/DR; clone CR3/43), and the polymer-based HRP-conjugated anti-mouse EnVision+ secondary were purchased from Dako (Carpinteria, CA). Heat-inactivated pooled human serum was purchased from Innovative Biologics (Herndon, VA). Dulbecco’s Modification of Eagle’s Medium (DMEM) was purchased from Corning Life Sciences (Tewksbury, MA).

Sillman B., Bade A.N., Dash P.K., Bhargavan B., Kocher T., Mathews S., Su H., Kanmogne G.D., Poluektova L.Y., Gorantla S., McMillan J., Gautam N., Alnouti Y., Edagwa B, & Gendelman H.E. (2018). Creation of a long-acting nanoformulated dolutegravir. Nature Communications, 9, 443.

+ Open protocol

+ Expand

Immunophenotyping of Mesenchymal Stem Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

The immunophenotype characterization of the MSCs was conducted using a previously described protocol [1 (link)]. Conjugated monoclonal antibodies against PE-Cy™5 mouse anti-human CD90, PE-Cy™7 mouse anti-human CD73, APC mouse anti-human CD13, FITC mouse anti-human HLA-ABC, APC mouse anti-human CD34, FITC mouse anti-human CD31, PE mouse anti-human CD45, PE mouse anti-human CD14 (BD Biosciences, San Diego, CA, USA), eFluor™450 mouse anti-human CD105, PE mouse anti-human CD29 (eBioscience, San Diego, CA, USA), PE/Cyanine7 mouse anti-human HLA-DR and PE/Cyanine7 mouse anti-human CD10 (BioLegend, San Diego, CA, USA) were used.

Cortés-Morales V.A., Chávez-Sánchez L., Rocha-Zavaleta L., Espíndola-Garibay S., Monroy-García A., Castro-Manrreza M.E., Fajardo-Orduña G.R., Apresa-García T., Gutiérrez-de la Barrera M., Mayani H, & Montesinos J.J. (2023). Mesenchymal Stem/Stromal Cells Derived from Cervical Cancer Promote M2 Macrophage Polarization. Cells, 12(7), 1047.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!