All cells were stained for viability using the Zombie Fixable Viability Kit (Biolegend), incubated with anti-CD16/32 Fc-block (BioXcell), and stained with the indicated antibodies: CD19 APC, CD3 APC-H7 or BV450, CD4 AF700, CD8 V500, CCR7 PE-CF594, CD69 BV785, CCR5 PE, CD103 FITC, (BD Biosciences) CD38 PE-Cy7, CD11c BV711, CD14 BV650, CD45RA BV605 (Biolegend). Stained samples were run on an LSRII flow cytometer, data acquired using FACS DIVA software (BD Biosciences) and analyzed using FlowJo software (TreeStar, Inc., Ashland, Oregon).
Cd103 fitc
Manufactured by BD
Sourced in United States
The CD103 FITC is a fluorescently labeled antibody that binds to the CD103 surface marker. CD103 is a type I transmembrane glycoprotein that is expressed on the surface of various immune cells, including intraepithelial lymphocytes, dendritic cells, and a subset of regulatory T cells. The FITC (fluorescein isothiocyanate) fluorescent label allows for the detection and analysis of CD103-expressing cells using flow cytometry or other fluorescence-based techniques.
Automatically generated - may contain errors
Lab products found in correlation
Facsdiva software, by BD (2 mentions)
Lsrii flow cytometer, by BD (1 mentions)
Cd19 apc, by BD (1 mentions)
Cd45ra bv605, by BioLegend (1 mentions)
Ccr7 pe cf594, by BD (1 mentions)
Ccr5 pe, by BD (1 mentions)
Cd8 v500, by BD (1 mentions)
Cd38 pe cy7, by BioLegend (1 mentions)
Zombie fixable viability kit, by BioLegend (1 mentions)
Cd4 af700, by BD (1 mentions)
Cd11c bv711, by BioLegend (1 mentions)
Cd14 bv650, by BioLegend (1 mentions)
Anti cd16 32 fc block, by BioXCell (1 mentions)
Facscalibur, by BD (1 mentions)
Cd4 fitc, by BioLegend (1 mentions)
Pe anti human cd25, by BD (1 mentions)
Cd40 fitc, by BioLegend (1 mentions)
Cd86 pe, by R&D Systems (1 mentions)
Ccr7 pe, by R&D Systems (1 mentions)
Hla dq fitc, by BD (1 mentions)
7 protocols using cd103 fitc
1
Comprehensive Immunophenotyping by Flow Cytometry
2
Phenotyping of Activated Monocyte-Derived Dendritic Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Phenotyping of resting and activated moDCs was performed by flow cytometry using anti-human CD1d-phycoerythrin (PE), CD103-FITC, HLA-DQ-FITC, PD-L1-PE (BD Biosciences, Franklin Lakes, NJ, USA), CD1a-allophycocyanin (APC), CD40-FITC (BioLegend, San Diego, CA, USA), CX3CR1-PE, CD80-FITC, CD83-FITC, CD86-PE, DC-SIGN-FITC, CCR7-PE, CD14-PE (R&D Systems, Minneapolis, MN, USA), B7RP1 (ICOSL)-PE (EBiosciences, Santa Clara, CA, USA), and isotype-matched control antibodies. The ratio of regulatory T-lymphocytes was measured by flow cytometry using anti-human CD25-PE (BD Pharmingen), CD4-FITC (BioLegend), FoxP3-APC (R&D Systems), and anti-IL-10-AlexaFluor488 (BioLegend). The viability of moDCs was determined with 2 μg/ml 7-amino-actinomycin D (LKT Laboratories Inc., St. Paul, MN, USA) dye followed by a 24-h activation period with live bacteria or LPS. Fluorescence intensities were measured by FACSCalibur (BD Biosciences), and data were analyzed by the FlowJo software (Tree Star, Ashland, OR, USA).
3
Multiparametric Flow Cytometry Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All flow cytometry reagents were from Biolegend unless otherwise stated. Data was acquired using a LSR-II flow cytometer (BD Biosciences) and analysed using FACS DIVA software (BD Biosciences); Fluorescence Minus One (FMO) controls established gating strategies. Figures were generated using FlowJo software (Tree Star Inc.). Immune cells were further purified by separation by density gradient centrifugation over lymphoprep (Axis Shield Diagnostics) then cells were washed twice with PBS/2% FCS and consecutive centrifugation of 800 × g and 200 × g for 10 minutes.
For flow cytometry, cells were incubated with antibodies for 20 minutes at 4 °C, washed ×2 in PBS/2% FCS and fixed in Fluorofix prior to data acquisition. Antibodies used were CD4-FITC(OKT4), CD8a-PE(HIT8a), CD3-PeCy5(UCHT1), CD56-PeCy7(BD Biosciences; B159), CD16-APCCy7(BD Biosciences; 3G8), CD45-APC(HI30), CD103-FITC(BER-ACT8), CD69-PE(FN90), CD8a-PECy7(RPA-T8), CD45RO-APC-Cy7(UCHL1), CD127-AlexaFluor647(A019D5), PD-1(CD279)-APC(EH12.2H7).
For flow cytometry, cells were incubated with antibodies for 20 minutes at 4 °C, washed ×2 in PBS/2% FCS and fixed in Fluorofix prior to data acquisition. Antibodies used were CD4-FITC(OKT4), CD8a-PE(HIT8a), CD3-PeCy5(UCHT1), CD56-PeCy7(BD Biosciences; B159), CD16-APCCy7(BD Biosciences; 3G8), CD45-APC(HI30), CD103-FITC(BER-ACT8), CD69-PE(FN90), CD8a-PECy7(RPA-T8), CD45RO-APC-Cy7(UCHL1), CD127-AlexaFluor647(A019D5), PD-1(CD279)-APC(EH12.2H7).
4
Isolation and Flow Cytometry of Dendritic Cell Subsets
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DCs were isolated from lymphoid organs as described previously (9 (link)). Briefly, single cell suspensions were prepared from pooled skin-draining lymph nodes and enriched for cDCs using antibody depletion and magnetic bead enrichment. Cells were incubated with anti-mouse monoclonal antibodies CD11c Brilliant Violet (BV) 421, MHC II APC-Cy7, CD8α PE-CF594, XCR1 PE, CD103 FITC, and CD326 PE-Cy7 purchased from BD Biosciences (Franklin Lakes, NJ, USA) or BioLegend (San Diego,CA, USA) to identify DC subsets. DCs were sorted from Propidium Iodide negative events using a FACSAriaIII cell sorter (BD Biosciences). During sorting, DCs were collected in buffer containing FCS (Sigma-Aldrich, St. Louis, MO, USA) and EDTA (Sigma-Aldrich) and kept on ice at all times before being recovered by centrifugation and stored in Trizol reagent (Invitrogen, Carlsbad, CA, USA).
5
Mouse Immune Cells Isolation Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Hank’s Balanced Salts Solution was from Invitrogen (Carlsbad, CA). ELISA kits were from eBioscience (San Diego, CA) (IL1α, IL1β, TNFα, IL6), from Immunology Consultants Laboratory Inc. (Portland, OR) (albumin), from R&D Systems (Minneapolis, MN) (CCL20) and from PeproTech (Rocky Hill, NJ) (CXCL1, CSF2, CSF3). Antibodies for flow cytometry analysis were from eBioscience (San Diego, CA) (anti-mouse CD45 eF450, CD11c PE, CD24 FITC, CD11b APC, MHC-II I-A/E PerCP-eF710, CD103 FITC) and from BD Biosciences (San Jose, CA) (anti-mouse Siglec F-PE and Ly6G (clone 1A8)-PE). QuickIII staining kit for cytospins was obtained from Astral Diagnostics, NJ. Butylated hydroxytoluene and indomethacin were from Fisher Scientific. Percoll, collagenase XI, Trypsin inhibitor, DNase I, RBC lysis solution were from Sigma-Aldrich (St. Louis, MO). Nylon cell strainers (70 μm) were from BD Biosciences (San Jose, CA). Qiasol lysis reagent, RNeasy Mini Kits and Mouse Cytokines & Chemokines RT2 Profiler PCR Array were from Qiagen (Valencia, CA). Paraformaldehyde was from Electron Microscopy Sciences (Hatfield, PA). XTT Cell Viability Kit was from Cell signaling.
6
Multiparametric Flow Cytometry of Tumor and Liver Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tumour and liver samples were enzymatically digested (0.25 mg/mL liberase (Sigma) and 1 mg/mL DNAse (Sigma) in DMEM) at 37 °C for 40 min. Cell suspensions were filtered (100 μm cell filters, Corning), treated with red blood cell lysis buffer for 2 min at 37 °C, washed, and resuspended at 1 × 107 cells/mL in cold FACS wash buffer (10% bovine serum albumin/5 mM EDTA/0.01% sodium azide in PBS pH 7). Briefly, 1 × 106 cell aliquots were stained with either Cocktail 1: CD45-BV510, CD4-FITC, CD3-PE, CD44-PerCP, CD8-PECy7, PD-1-APC, CD69-APC-Cy7 (BD, Biosciences) and viability dye DAPI, Cocktail 2: CD45-BV510, CD103-FITC, CD3-PE, CD8-PECy7, PD-1-APC, CD4-APC-Cy7 (BD, Biosciences) and DAPI, or Cocktail 3: CD45-BV510, F4/80-FITC, Ly6C-PE, PD-L1-PECy7, CD11b-APC-Cy7 (BD, Biosciences), and DAPI. Samples were run on the Canto II (BD Biosciences) and analysed using FlowJo™ software.
7
In Vivo Labeling of Circulating T 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!