The CD45- fraction left after CD45+purification was used to sort FAP+ CAFs from the tumors. CD45- cell suspensions were stained with a live/dead marker and FAP-Biotin (R&D Systems, Minneapolis, MN, USA) followed by secondary staining with streptavidin as described above. Up to 3 x 105 cells were sorted into PBS. The samples were sorted using a FACS Aria I cell sorter (BD Biosciences, San Jose, CA, USA). Immediately after sorting, cells were processed for RNA isolation.
Fap biotin
Manufactured by R&D Systems
Sourced in United States
FAP-Biotin is a lab reagent used to detect and quantify the presence of Fibroblast Activation Protein (FAP) in biological samples. It is a biotinylated antibody that specifically binds to the FAP protein, allowing for its detection and measurement through various analytical techniques.
Automatically generated - may contain errors
Lab products found in correlation
Facsaria 1 cell sorter, by BD (1 mentions)
Streptavidin apc, by Thermo Fisher Scientific (1 mentions)
Pdpn apc, by Thermo Fisher Scientific (1 mentions)
Cd26 percp cy5, by Thermo Fisher Scientific (1 mentions)
α sma cy3, by Merck Group (1 mentions)
Cd8 bv605, by BD (1 mentions)
Ly6c percpcy5, by R&D Systems (1 mentions)
Pd l1 apc, by BD (1 mentions)
Cd31 fitc, by R&D Systems (1 mentions)
F4 80 fitc, by R&D Systems (1 mentions)
Zombie aqua, by BioLegend (1 mentions)
Facscanto 2, by BD (1 mentions)
Cd45 fitc, by BD (1 mentions)
Cd3 af700, by BD (1 mentions)
Cd4 bv421, by BD (1 mentions)
2 protocols using fap biotin
1
Isolation of FAP+ Cancer-Associated Fibroblasts
2
Comprehensive Immune Cell Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The following antibodies/dies (purchased from BioLegend unless otherwise stated) were used for flow cytometry: CD45-PE-Cy7, CD31-FITC, FAP-biotin (R&D system), streptavidin-APC, CD90-BV605, PDPN-APC, Ly6C-AF700, CD26-PerCP-Cy5.5 (eBioscience), αSMA-Cy3 (Sigma), CD11b-PE-Cy7, CD11b-Pacific Blue, F4/80-APC, F4/80-FITC, MR-PE, MR-PE-Cy7, Ly6C-PerCP-Cy5.5, Ly6G-APC-Cy7, Arg1-PE (R&D system), PDL1-APC (BD Biosciences), CD45-FITC, CD3-AF700, CD4-BV421, CD8-BV605 (BD Biosciences), CD25-PE-Cy7, FoxP3-APC and carboxyfluorescein succinimidyl ester (CFSE, Sigma Aldrich). Viability was assessed with Zombie Aqua (BioLegend). Samples were Fc receptor-blocked using mouse FcR blocking reagent (1:10; Miltinyi Biotec). For intracellular staining, samples were fixated and permeabilized with eBioscience Fixation/Permeabilization Concentrate, Diluent and 10X Buffer (Invitrogen). Data were acquired on FACSCanto II (BD Biosciences) or ACEA NovoCyte Quanteon (Agilent) and analyzed with FlowJo V.10.6.1 (Tree Star). Gating strategies can be found in online supplemental figures 8–13 .
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