For flow cytometry analysis cells were stained with CD14-PE-Cy7 (1:20, BD Bioscience, Breda, the Netherland), TLR4-APC (1:10, ebioscience, San Diego, USA) or left untreated. Cell numbers were quantified using the BD FACS Cantoll flowcytometer, histograms were generated using flowjo software version 7.6.2 (Treestar Inc, Ashland-OR, USA.
Tlr4 apc
Manufactured by Thermo Fisher Scientific
Sourced in United States
The TLR4-APC is a lab equipment product manufactured by Thermo Fisher Scientific. It is a fluorochrome-conjugated monoclonal antibody that binds to the Toll-like receptor 4 (TLR4) protein on the surface of cells. The core function of this product is to facilitate the detection and analysis of TLR4 expression in various cell types using flow cytometry or other immunoassay techniques.
Automatically generated - may contain errors
Lab products found in correlation
Facs cantoll flow cytometer, by BD (1 mentions)
Cd14 pe cy7, by BD (1 mentions)
Tlr2 pe, by Thermo Fisher Scientific (1 mentions)
Cd28 percp cy5.5, by Thermo Fisher Scientific (1 mentions)
Cd45ra pe cy7, by Thermo Fisher Scientific (1 mentions)
Facsaria, by BD (1 mentions)
Lps from e coli, by Merck Group (1 mentions)
3 protocols using tlr4 apc
1
Flow Cytometry Analysis of CD14 and TLR4
2
Characterization of Naive CD4 T Cells
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We analyzed naive CD4 T-cell surface markers before and after in vitro culture of isolated naive CD4 T cells in flat-bottom plates (250 × 105 T cells/well), with and without CD3/CD28 Dynabeads human T-activator CD3/CD28 (Invitrogen, Grand Island, NY), under optimal stimulation conditions (25 µL beads/106 naive CD4 T cells) for 24 and 48 hours in complete RPMI 1640 medium containing 10% heat-inactivated fetal bovine serum, 2 mM l -glutamine, 100 U/mL penicillin/streptomycin in 5% CO2 at 37°C. We used a FACSAria (Becton Dickinson, San Diego, CA) to analyze T-cell aliquots stained with the following antibodies: CD45RA PE-Cy7, TLR2 PE, TLR4 APC, CD28 PerCP-Cy 5.5 (eBioscience, San Diego, CA), and CCR1 Alexa Fluor 488 (R&D Systems, Minneapolis, MN). We defined CD4+ CD45RA+ CD45RO− CD62L+ T cells as naive. Purity was >90% in all samples, with the majority >95%.
3
LPS Stimulation of LDLr-Deficient Mast Cells
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LDLr−/− and LDLr−/−/RP105−/− BMMCs were seeded at a density of 106 cells per well (n = 3), after which they were stimulated with 1 ng/mL, 10 ng/mL or 100 ng/mL LPS (from E. Coli, Sigma-Aldrich, St. Louis, MO, USA). After 4 hours and 24 hours activation, cells were spun down (1500 rpm, 5 minutes) and the supernatant was collected for further analysis. ELISAs were performed according to manufacturer’s protocol. Expression of the surface markers RP105 (RP105-PE, Ebioscience, 12-1801-81) and TLR4 (TLR4-APC, Ebioscience, 12-9924-82) on control LDLr−/− mast cells was determined by means of FACS analysis.
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