RPMI 1640 was purchased from Life Technologies Corporation (Grand Island, NY, USA), containing 4.5 g/L D-Glucose, L-Glutamine and 110 mg/L Sodium Pyruvate. GM-CSF, IL-4 and TNF-α were purchased from Peprotech Asia (Revohot, Israel). Lipopolysaccharide (LPS) was purchased from Sigma Aldrich (Steinheim, Sweden). D-luciferin was purchased from Promega Corporation (Madison, WI, USA). The following antibodies were used for FACS analysis: fluorescein isothiocyanate (FITC)-conjugated anti-CD86, FITC-conjugated anti-I-A[b], FITC-conjugated anti-H-2Kb, Phycoerythrin (PE)-conjugated anti-CCR7, PE-conjugated anti-mouse CD40, PE-conjugated anti-CD11c and FITC conjugated anti-CD11c were purchased from BD PharMingen (SanDiego, CA). PE-conjugated anti-H-2Kb (SIINFEKL) was purchased from BioLegend (San Diego, CA). PE-conjugated anti-CCR1, PE-conjugated anti-CCR2, PE-conjugated anti-CXCR1 and PE-conjugated anti-CCR7 were purchased from R&D Systems (Minneapolis, MN, USA). All the isotype control antibodies were purchased from the same company as the detecting antibodies.
Fitc conjugated anti cd11c
Manufactured by BD
Sourced in United States
FITC-conjugated anti-CD11c is a monoclonal antibody that binds to the CD11c cell surface antigen. CD11c is expressed on the surface of dendritic cells, macrophages, and some lymphocytes. The antibody is conjugated with the fluorescent dye FITC (fluorescein isothiocyanate), which allows for the identification and enumeration of CD11c-positive cells using flow cytometry or other fluorescence-based techniques.
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Lab products found in correlation
Pe conjugated anti foxp3, by BD (4 mentions)
Bv605 conjugated anti cd11b, by BD (4 mentions)
Bv450 conjugated anti cd4, by BD (4 mentions)
Fitc conjugated cd25, by BD (4 mentions)
Fitc conjugated anti cd86, by BD (2 mentions)
Fitc conjugated anti cd8, by BD (2 mentions)
Apc conjugated anti cd11b, by BD (2 mentions)
Lsrii flow cytometer, by BD (2 mentions)
Pe conjugated anti ccr1, by R&D Systems (1 mentions)
Pe conjugated anti ccr2, by R&D Systems (1 mentions)
D glucose, by Thermo Fisher Scientific (1 mentions)
Pe conjugated anti cd11c, by BD (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Lipopolysaccharide lps, by Merck Group (1 mentions)
D luciferin, by Promega (1 mentions)
Apc conjugated anti cd3, by BD (1 mentions)
Pe conjugated anti cd4, by BD (1 mentions)
Macsquant flow cytometer, by Miltenyi Biotec (1 mentions)
Apc conjugated anti cd19, by BD (1 mentions)
Apc conjugated anti ifn γ, by BD (1 mentions)
10 protocols using fitc conjugated anti cd11c
1
Murine Dendritic Cell Differentiation
2
Intracellular Cytokine Staining in Lung Cells
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After lung cell purification, intracytoplasmic cytokine staining was performed as previously described [26 (link), 27 (link)]. The cells were stained for cell surface markers with PE-conjugated anti-CD4 (BD Biosciences, Franklin Lakes, NJ, USA), FITC-conjugated anti-CD8 (BD Biosciences), or APC-conjugated anti-CD3 (BD Biosciences) and then analyzed using a MACS Quant flow cytometer (Miltenyi Biotec, Auburn, CA, USA) with FlowJo software (TreeStar, Ashland, OR, USA). The numbers of cytokine-producing CD4+ or CD8+ T cells per lung were calculated from the percentages of cytokine-producing cells and the numbers of CD4+ or CD8+ T cells isolated from the lung. The cells were also stained for cell surface markers with APC-conjugated anti-CD11b (BD Biosciences) and FITC-conjugated anti-CD11c (BD Biosciences).
3
Murine LAIR-1 Antibody Characterization
4
Flow Cytometric Analysis of BMDC Phenotype
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BMDCs and BMDC/NF cells were stained with PE-conjugated anti-CD54, anti-CD86, anti–H-2Kb (MHC class I,) or anti–I-A/I-E (MHC class II); FITC-conjugated anti-CD11c; or APC-Cy7–conjugated anti-CD197 (CCR-7) antibodies (BD Biosciences) at 4°C for 30 minutes. After washing with PBS, the cells were analyzed using a BD Accuri C6 flow cytometer. Isotype-matched monoclonal antibodies were used as controls. The data were analyzed using the FlowJo analysis software (FlowJo, Ashland, OR).
5
Isolation and Characterization of Mouse Uterine and Placental Mononuclear Cells
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Mononuclear cells from mouse uterine and placenta were prepared as previously described (Liu X. et al., 2014 (link)). Briefly, mice were sacrificed on Gd 14. The uterus and placenta were excised with surgical cuts. The pregnancy outcome was reflected by the total number of fetuses, fetal size, stillbirth, absorptive fetus, and hemorrhagic appearance. The uterus and placentas were washed with sterile PBS, minced into small pieces. Dispersed cells were collected by filtration through a 48 μm pore size stainless steel mesh. Thereafter, the mononuclear cells were obtained by density-gradient centrifugation and washed twice in cold PBS. The following fluorochrome-conjugated, mouse-specific mAbs were used in the assays: FITC-conjugated anti-CD11c (BD Biosciences, United States), Percp-cy5.5-conjugated anti-CD8a mAb (BD Biosciences, United States), PE-conjugated anti-CD80 (BD Biosciences, United States), PE-conjugated anti-CD86 (BD Biosciences, United States), PE-conjugated anti-I-Ad/I-Ed (MHC II) (BD Biosciences, United States), and PE-conjugated anti-LILRB4 (Biolegend, United States). Cells were incubated for 30 min at 4°C in the dark according to the manufacturer’s instructions. Subsequently, the cells were washed with cold PBS and analyzed using BD FACSAria flow cytometry and BD FACSDiva 7.0 software (Becton Dickinson, United States).
6
FACS-based Immune Cell Profiling
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Single-cell suspensions were stained in fluorescence-activated cell sorting (FACS) buffer at 4 °C for 30 min. We analyzed samples using an LSRII flow cytometer (BD Pharmingen, San Diego, CA). The following antibodies against mouse antigens were purchased from BD Pharmingen (San Diego, CA): BV605-conjugated anti-CD11b, FITC-conjugated anti-CD11c, BV450-conjugated anti-CD4, FITC-conjugated CD25, and PE-conjugated anti-Foxp3.
7
Flow Cytometric Analysis of Dendritic Cell Maturation
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The iDC and mDC were washed with phosphate-buffered saline (PBS), aliquoted into fractions (5 × 105 cells/100 μl), and then stained for 30 min in the dark at room temperature with a final concentration of 5 μg/ml of the following antibodies purchased from BD Biosciences (San Jose, CA, USA): fluorescein isothiocyanate (FITC)-conjugated anti-CD11c (553801), FITC-conjugated anti-CD40 (553790), FITC-conjugated anti-CD80 (553768), and FITC-conjugated anti-CD86 (553691). As for the negative or no-antibody control, the cells were also stained with corresponding FITC-conjugated isotype-matched control antibodies or remained unstained. After staining, the cells were washed with PBS twice and analyzed by a BD LSRII flow cytometer (BD Biosciences). Data analysis was performed using FlowJo software (Tree Star, San Carlos, CA, USA). The cells positive for FITC-CD11c were considered as DC that had successfully differentiated from bone marrow cells. The cells positive for FITC-CD40, FITC-CD80, and FITC-CD86 were considered as DC that had undergone successful maturation. For concurrent analysis of these molecules, the mDC were stained with phycoerythrin-indotricarbocyanine (PE-Cy7)-conjugated anti-CD11c (558079; BD Biosciences), together with the FITC-conjugated anti-CD40, FITC-conjugated anti-CD80, and FITC-conjugated anti-CD86, respectively.
8
Multicolor Flow Cytometry of Immune Cells
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Single-cell suspensions were stained in uorescence-activated cell sorting (FACS) buffer at 4°C for 30 min. We analyzed samples using an LSRII ow cytometer (BD Pharmingen, San Diego, CA). The following antibodies against mouse antigens were purchased from BD Pharmingen (San Diego, CA): BV605conjugated anti-CD11b, FITC-conjugated anti-CD11c, BV450-conjugated anti-CD4, FITC-conjugated CD25, and PE-conjugated anti-Foxp3.
9
Flow Cytometry Analysis of Immune Cells
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Single-cell suspensions were stained in FACS buffer at 4 °C for 30 min. We analyzed Samples using an LSRII (BD Pharmingen, San Diego, CA). The following antibodies against mouse antigens were purchased from BD Pharmingen (San Diego, CA): BV605-conjugated anti-CD11b, FITC-conjugated anti-CD11c, BV450-conjugated anti-CD4, FITC-conjugated CD25, and PE-conjugated anti-Foxp3.
10
Flow Cytometry of Immune Cells
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Single-cell suspensions were stained in fluorescence-activated cell sorting (FACS) buffer at 4°C for 30 min. We analyzed samples using an LSRII flow cytometer (BD Pharmingen, San Diego, CA). The following antibodies against mouse antigens were purchased from BD Pharmingen (San Diego, CA): BV605-conjugated anti-CD11b, FITC-conjugated anti-CD11c, BV450-conjugated anti-CD4, FITC-conjugated CD25, and PE-conjugated anti-Foxp3.
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