CD3-percp-cy5.5 (Catalogue #45-0036-42), CD8-APC (Catalogue #17-0086-42), CD8-FITC (Catalogue #11-0086-42), CD14-APC (Catalogue #17-0149-42), CD56-FITC (Catalogue #4278380), CD16-PerCP-eFluor™710 (Catalogue #46-0168-42), CD11b-PerCP-eFluor™710 (Catalogue #46-0110-80), IFN-γ-PE (Catalogue #12-7319-42), TLR2-FITC (Catalogue #11-9922-41) and Mouse IgG1-PE (Catalogue #12-4714-81) were all bought from eBioscience. HLA-DR-PE (Catalogue #555812), IL-10-APC (Catalogue #554707), CXCR3-APC (Catalogue #550967) and IFN-γ-FITC (Catalogue #561053), Rat IgG2a-APC (Catalogue #554690) and mouse IgG-APC (Catalogue #5065947) were purchased from BD Biosciences. CD16-FITC (Catalogue #302006), HLA-DR-APC (Catalogue #307609), CD3-FITC (Catalogue #317306) and TLR4-APC (Catalogue #312815) were purchased from Biolegend. EP2-PE (Catalogue #10477) and Rabbit IgG-PE (Catalogue D5-1610) were purchased from Cayman Chemical.
Tlr2 fitc
Manufactured by Thermo Fisher Scientific
Sourced in United States
TLR2-FITC is a fluorescently-labeled antibody that binds to the Toll-like receptor 2 (TLR2) protein. TLR2 is a cell surface receptor that plays a role in the innate immune response.
Automatically generated - may contain errors
Lab products found in correlation
Tlr4 pe, by Thermo Fisher Scientific (3 mentions)
Cellquest software, by BD (3 mentions)
Hla dr pe, by BD (2 mentions)
Cd80 fitc, by BD (2 mentions)
Cd86 pe, by BD (2 mentions)
Facs buffer, by BD (2 mentions)
Cd14 percp, by BD (2 mentions)
Facscalibur, by BD (2 mentions)
Cd40 pe, by BD (2 mentions)
Cd8 fitc, by Thermo Fisher Scientific (1 mentions)
Cd14 apc, by Thermo Fisher Scientific (1 mentions)
Tlr4 apc, by BioLegend (1 mentions)
Ifn γ fitc, by BD (1 mentions)
Cxcr3 apc, by BD (1 mentions)
Cd8 apc, by Thermo Fisher Scientific (1 mentions)
Cd3 percp cy5, by Thermo Fisher Scientific (1 mentions)
Hla dr apc, by BioLegend (1 mentions)
Cd3 fitc, by BioLegend (1 mentions)
Mouse igg1 pe, by Thermo Fisher Scientific (1 mentions)
Cd56 fitc, by Thermo Fisher Scientific (1 mentions)
4 protocols using tlr2 fitc
1
Multiparameter Immune Cell Profiling
2
Monocyte Surface Marker Expression by Flow Cytometry
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Cells were incubated with the following monoclonal antibodies: CD14-PerCP (BD Pharmingen, USA), CD16-FITC (BD Pharmingen), CD80-FITC (BD Pharmingen), CD86-PE (BD Pharmingen), CD40-PE (BD Pharmingen), TLR2-FITC (eBioscience, USA), and TLR4-PE (eBioscience). In isotype controls cells were stained with IgG1 conjugated with the respective fluorochromes (BD Pharmingen, eBioscience). After 30 min of incubation at 4°C, cells were washed twice in FACS buffer (Becton Dickinson, USA). The expression of monocytes surface markers was measured by flow cytometry (FACSCalibur, Becton Dickinson) and analyzed by Cell Quest software (Becton Dickinson). The results were based on analysis of at least 100,000 cells and were shown as the percentage of positively labeled cells. Moreover, the mean fluorescence intensity (MFI) values of gated monocytes positive for individual markers were determined.
3
Flow Cytometry Analysis of Immune Cells
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Cellular population in BALF were washed and resuspended at a concentration of 0.5×106 to 1×106/50 µl FA buffer (Difco) plus 0.1% NaN3, and Fc receptors were blocked by the addition of unlabeled anti-CD16/32(Fc blocking, eBioscience Inc.). After Fc receptor blocking, immunostaining for cell surface molecules was performed for 30 min at 4°C. Cells were washed twice with FA buffer, resuspended in 200 µl of 4% formalin (Sigma). A minimum of 10000 events were acquired on a FACSCanto flow cytometer (BD Pharmingen) using CellQuest software (BD Pharmingen). The data acquired were analyzed with FlowJo software (Tree Star, Stanford, CA). Fluorochrome-conjugated antibodies directed against the following antigens were obtained: CD45-PE, Gr-1-FITC, CD11c-APC, TLR2-FITC, TLR4-PE, (eBioscience Inc.), and Dectin-1-PE (R&D). Numbers of relevant cell types were determined by combining flow cytometry data (percentage of a given cell type).
4
Phenotypic Characterization of CD1c+ DCs
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Cells were incubated with appropriate monoclonal anti bodies: CD11c-APCw (BD Pharmingen), CD1c-FITC (eBioscience), CD83-PE (BD Pharmingen), CD80-FITC (BD Pharmingen), CD86-PE (BD Pharmingen), CD40-PE (BD Pharmingen), PD-L1-FITC (BD Pharmingen), PD-L2-PE (BD Pharmingen), HLA-DR-PE (BD Pharmingen), CCR7-APC-e Fluor (eBioscience), TLR2-FITC (eBioscience), TLR4-PE (eBioscience), CD64-FITC (BD Pharmingen), and DEC-205-PE (BD Pharmingen). In addition, CD14-PerCP (BD Pharmingen) monoclonal antibody was used to confirm the differentiation of DCs. Isotype controls were cells stained with IgG1 conjugated with the respective fluorochromes (BD Pharmingen, eBioscience). After 30 min of incubation at 4°C, cells were washed twice in FACS buffer (Becton Dickinson). The expression of individual markers on gated CD1c+ DCs was determined by flow cytometry (FACSCalibur, Becton Dickinson) and analyzed by Cell Quest software (Becton Dickinson). The results were based on analysis of at least 100,000 cells and were shown as the percentage of positively labeled cells. In addition, the mean channel fluorescence values of gated DCs positive for individual markers were determined.
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