BAL and lung cells were treated with RBC lysing buffer (Sigma). Lungs were dissociated using a Lung Dissociation Kit (Miltenyi Biotec) and Gentle MACS (Miltenyi Biotec). LIVE/DEAD cells were stained with Fixable Aqua Dead Cell Staining Kit (Thermo Fisher Scientific), and after Fc block with the 2.4G2 mAb (eBioscience), cells were stained with the following Abs: DR3-PE (4C12; BioLegend), SiglecF (E50-2440; BD Biosciences), CD11b (M1/70; BD Biosciences), CD11c (HL3; BD Biosciences), Ly6G (1A8; BioLegend), ST2-PE (101001PE, MD), CD90.2 (30-H12; BioLegend). For lineage markers for ILC2 staining, the following Abs were used: CD3-FITC (145-2C11; eBioscience), CD4-FITC (GK1.5; eBioscience), CD8- FITC (5H-10-1; BioLegend), CD19-FITC (1D3; BioLegend), NK1.1-FITC (PK136; BioLegend), CD11b-FITC (M1/70; BioLegend), CD11c-FITC (HL3; BD Biosciences), and Gr1-FITC (RB6-8C5; BioLegend). Flow cytometry analysis was performed on a Fortessa (BD Biosciences), and data were analyzed using FlowJo Software (version 10; FlowJo, Ashland, OR). Live CD45+ lung immune cells were separated into T cells (CD3+, CD90.2+), macrophages (CD11b+, CD11c+ SiglecF+), DCs (CD11c+ MHC class II+), neutrophils (GR1+, CD11b+ SigF-), eosinophils (Ly6C+, SigF+, CD11c-) and ILC2 (Lin- Thy1.2+ ST2+Sca1+).
Cd11c fitc
Manufactured by BD
Sourced in United States
CD11c-FITC is a fluorescently-labeled antibody that binds to the CD11c cell surface antigen, which is expressed on dendritic cells and some other immune cells. It can be used for the identification and analysis of these cell populations by flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Facscalibur, by BD (6 mentions)
Cd4 apc, by BD (5 mentions)
Siglecf pe, by BD (5 mentions)
F4 80 pe, by BD (4 mentions)
Cd86 pe, by BD (4 mentions)
Facscanto 2, by BD (4 mentions)
Cd11b fitc, by BD (4 mentions)
Cd3 fitc, by BD (4 mentions)
Cd11b apc, by BD (4 mentions)
Cd44 fitc, by BD (3 mentions)
Cd11b pe cy7, by BD (3 mentions)
Cd4 apc cy7, by BD (3 mentions)
Cd8 apc, by BD (3 mentions)
Cd4 pe, by BD (3 mentions)
Cd11c pe, by BD (3 mentions)
Cd8 pe, by BD (3 mentions)
Foxp3 pe, by BD (3 mentions)
B220 apc, by BD (2 mentions)
Foxp3 pe, by Thermo Fisher Scientific (2 mentions)
Cd4 pe cy7, by BD (2 mentions)
37 protocols using cd11c fitc
1
Comprehensive Lung Immune Cell Profiling
2
Immunophenotyping of Murine Macrophages
3
Multiparameter Flow Cytometry Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For cell surface staining, empirically determined dilutions of primary mAbs were used to stain single cell suspensions on ice for 20 min in FACS buffer (PBS with 2% FBS, 0.1% NaN3, and 1 µM EDTA). All mAbs were purchased from BioLegend unless otherwise indicated. The following mAb clones were used: B220-PE-Cy7, CD4-PE-Cy7, CD8α-PerCP-Cy5.5, CD11b-PE-Cy7, CD11c-FITC, CD25–Alexa Fluor 647, CD44-FITC, CD45.1-FITC, CD45.2-BV605, CD62L-PE-Cy7, CD103-biotin (followed by streptavidin-BV711; BD), Foxp3-PE (eBioscience), Helios-APC, I-A(b)-PE, Ki-67–Alexa Fluor 647, and TCR-β–Pacific blue. Dead cells were excluded using Fixable Viability Dye eFluor780 (eBioscience). Intracellular staining to detect Foxp3 expression was performed according to the manufacturer’s instructions, using the manufacturer’s protocol for Foxp3 staining in 96-well plates (eBioscience). Cells were analyzed using an LSR-II flow cytometer (BD) equipped with 405-, 488-, 552-, and 640-nm lasers. Flow cytometry data were processed using FlowJo version 9.7.5 software (Tree Star).
4
Comprehensive Immune Cell Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell surface marker analysis was performed using flow cytometry. Single-cell suspensions prepared from MLN, adipose tissue (AT) and liver were collected from mice at the times indicated. Cell surface markers were stained for 30 min at 4°C with rat anti-mouse CD3/CD19-CF594 (Clone:145-2C11,1D3) F4/80-APC (Clone: T45-2342), CD11c-FITC (Clone: HL3), CD301-pecy7 (Clone: LOM-14), CD64-PerCp-Cy5.5 (Clone: X45-5/7.1), CD11b-BV650 (Clone: M1/70), Ly6G-efluor700 (Clone: 1A8) and Siglec-F-PE (Clone: E50-2440) (BD Bioscience). All antibody incubations were performed at 4°C for 30 min (isotype controls were included). Data were acquired using a BD FACS Aria and analyzed using FlowJo software (Tree Star, Inc).
5
Multiparametric flow cytometry analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell surface molecules were analysed by a FACSCalibur® flow cytometer (Becton Dickinson). Cells were incubated for 10 min at 4°C with relevant or control antibodies in 100 µl total staining volume in PBS-BSA. After staining, the cells were washed and resuspended in 300 µl of PBS-BSA. The following antibodies were used for the staining of human monocytes, T cells and dendritic cells:, anti-CD1c-FITC [anti-BDCA-1] (Miltenyi), anti-CD11b-PE [Mac-1] (BD Pharmingen), anti-CD14-APC (BD Pharmingen), anti-CD40-FITC (BD Pharmingen), anti-CD45RA-APC (BD Pharmingen), anti-CD86-PE (eBioscience), anti-HLA-DR-CyChrome (BD Pharmingen). The following antibodies were used for the staining of murine dendritic cells and T cells: CD4-AF647, vα2-Bio, SA-PacificBlue, CD11c-FITC, CD86-PE, MHC II was measured by a combination of biotin-labeled I-Ab/SA-APC (all BD Pharmingen). To measure proliferation, T cells were labeled with the fluorescent dye CFSE (Molecular Probes).
6
Isolation and Analysis of Stromal Vascular Fraction
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The SVF was isolated from inguinal subcutaneous or epidydimal visceral fat depot of male FGF21KO and WT mice after rmFGF21 treatment or fat transplantation. The mice were anesthetized and the fat pads were removed and digested in 0.1% (w/v) collagenase type I (Invitrogen) for 30 min at 37 °C with gentle shaking. The digestion mixture was passed through a 100 mm cell strainer (BD Biosciences) and centrifuged at 800×g for 10 min at 4 °C. The SVF pellets were collected and washed twice, 1 × 105 freshly isolated cells were triple stained with F4/80-PE (Abcam, ab105156, clone CI:A3-1, 1:50), cd206-Alexa Fluor 647 (Biolegend, 141712, clone C068C2, 1:100) and cd11c-FITC (BD Biosciences, BD 557400, clone HL3, 1:100) on ice for 30 min in dark. Species-matched IgG was used as non-specific isotype controls. After washing, the cells were analyzed with LSR Fortessa Cell Analyzer (BD Biosciences).
7
Comprehensive Immune Cell Phenotyping
8
Tumor Dissociation and Immune Cell Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After cervical dislocation of mice, tumors were excised, minced and then incubated in an enzyme mixture (7.5 mg collagenase type I, 2.5 mg hyaluronidase type I-S (both Sigma-Aldrich, Germany) in 10 ml PBS) at 37°C for 1 h. After filtration of the cell suspension through a 100 μm nylon mesh erythrocytes were lyzed in 155 mM NH4Cl, 10 mM KHCO3, 0.1 mM Na2-EDTA (pH 7.4, 4°C, 8 min) and washed with PBS.
For FACS analysis cells were stained using the following anti-mouse antibodies: anti-CD4-FITC, CD8a-FITC, CD11b-FITC, CD11c-FITC, CD45-APC (all BD, Germany), CD25-PE (Miltenyi Biotec, Germany), as well as appropriate isotype controls. Cells were fixed in 4% (v/v) paraformaldehyde after staining. Analysis was performed on a BD FACSCanto II using the FACS Diva Software (both BD, Germany). Gates of immune cell subpopulations were defined by staining with respective isotype control antibodies and fractions are given as percentage of CD45+ immune cells.
For the IFN-γ-ELISpot 2 × 105 cells of the tumor were used and protocol was run according to the manufacturer’s specification (R&D Systems, Germany). The number of IFN-γ secreting cells was determined by counting the number of visible spots on the plate.
For FACS analysis cells were stained using the following anti-mouse antibodies: anti-CD4-FITC, CD8a-FITC, CD11b-FITC, CD11c-FITC, CD45-APC (all BD, Germany), CD25-PE (Miltenyi Biotec, Germany), as well as appropriate isotype controls. Cells were fixed in 4% (v/v) paraformaldehyde after staining. Analysis was performed on a BD FACSCanto II using the FACS Diva Software (both BD, Germany). Gates of immune cell subpopulations were defined by staining with respective isotype control antibodies and fractions are given as percentage of CD45+ immune cells.
For the IFN-γ-ELISpot 2 × 105 cells of the tumor were used and protocol was run according to the manufacturer’s specification (R&D Systems, Germany). The number of IFN-γ secreting cells was determined by counting the number of visible spots on the plate.
9
Cytokine and Flow Cytometry Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For cell culture and virus preparation, Dulbecco's Modified Eagle Medium (DMEM) and fetal bovine serum (FBS) were purchased from Life Technologies (Grand Island, NY, USA) and Lonza (Basel, Switzerland), respectively. To measure cytokine concentrations, cytometric bead array (CBA) mouse inflammation/Th1/Th2/Th17 cytokine kit, mouse IL-5 and IL-13 flex sets were purchased from BD Biosciences (San Diego, CA, USA). All reagents for flow cytometry including Golgi Plug, Cytofix/Cytoperm solution, anti-mouse CD16/32 (Mouse BD Fc BlockTM), CD4-APC-Cy7, CD44-FITC, IFN-γ-APC, IL-17-PE, IL-5-APC, CD11c-FITC, CD45-PerCP, Siglec-F-PE, and Ly6G-PE-Cy7 were purchased from BD Biosciences. For development of ELISA, horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG was purchased from Southern Biotechnology (Birmingham, AL, USA). HRP-conjugated goat anti-mouse IgG1 and IgG2a were purchased from Zymed Laboratories (San Francisco, CA, USA).
10
Characterization of Mesenchymal Stem Cells
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!