Phenotypically distinct muscle and tumor cell subsets were sorted according to protocols that were previously established to isolate functionally discrete subsets of myofiber-associated cells [10 (link), 11 (link), 1 (link)]. In brief, cells were suspended in HBSS supplemented with 2% FBS. Antibody staining was performed for 20 minutes on ice. The following primary and secondary antibodies were used: APC-CY7-conjugated anti-mouse CD11b (1 in 200, BD Pharmingen, 557657), APC-CY7-conjugated anti-mouse CD45 (1 in 200, eBioscience, 557659), APC-CY7-conjugated anti-mouse TER119 (1 in 200, BioLegend, 116223), APC-conjugated anti-mouse Sca1 (1 in 200, eBioscience, 17-5981-82), PE-conjugated anti-mouse/ rat CD29 (1 in 400, BioLegend, 102207), biotin rat anti-mouse CD184 (1 in 100, BD Pharmingen, 551968), PECY7-conjugated Streptavidin (1 in 200, eBioscience, 25-4317-82). Antibody staining was performed for 20 minutes on ice. Prior to FACS sorting, cells were suspended in 1µg/ml propidium iodide and 10µM calcein blue (Invitrogen) to identify viable cells (Pi−Ca+). Cells were sorted twice to maximize purity.
Biotin rat anti mouse cd184
Manufactured by BD
Biotin rat anti-mouse CD184 is a laboratory reagent used for the detection and analysis of CD184, also known as CXCR4, a chemokine receptor expressed on the surface of various cell types. This product is a biotin-conjugated monoclonal antibody raised in rats that specifically binds to the mouse CD184 antigen.
Automatically generated - may contain errors
Lab products found in correlation
Calcein blue, by Thermo Fisher Scientific (1 mentions)
Propidium iodide, by Thermo Fisher Scientific (1 mentions)
Apc cy7 conjugated anti mouse cd11b, by BD (1 mentions)
Apc cy7 conjugated anti mouse ter119, by BioLegend (1 mentions)
Pe cy7 conjugated streptavidin, by Thermo Fisher Scientific (1 mentions)
Pe cy7 anti mouse ly6a e sca 1 clone d7, by Thermo Fisher Scientific (1 mentions)
2 protocols using biotin rat anti mouse cd184
1
Isolation of Muscle and Tumor Cell Subsets
2
Satellite Cell Immunophenotyping Protocol
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For additional characterization of satellite cells, we stained for lineage negative; CXCR4 and Itgb1 double positive cells. PE-Cy7 anti-mouse TER-119 (clone TER-119), APC anti-mouse/rat CD29 (clone eBioHMb1-1), and PE-Cy7 anti-mouse Ly6A/E (Sca-1) (clone D7) from eBioscience (San Diego, CA); PE-Cy7 rat anti-mouse CD11b (clone M1/70), PE-Cy7 CD45, and Biotin rat anti-mouse CD184 from BD Biosciences antibodies were incubated with samples on ice for 1 h. Additionally, we stained for lineage negative; CD34, Itga7 double-positive cells. PE-Cy7 Rat anti-mouse CD11b (clone M1/70), PE-Cy7 anti-mouse Ly6A/E (Sca-1) (clone D7), and Biotin anti-mouse CD34 (clone RAM34) from eBioscience (San Diego, CA) and Itga7 PE (clone R2F2) from AbLab (Vancouver, B.C., Canada) antibodies were incubated for 1 h on ice. After this incubation, samples were washed and resuspended with FACS staining medium. PE or APC conjugated Streptavidin reagents, respectively, were incubated on ice for 20 min. Samples were washed and resuspended with FACS staining medium for FACS analysis.
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