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Mem 166

Manufactured by BioLegend
Sourced in United States

MEM-166 is a monoclonal antibody that recognizes the CD166 antigen. CD166 is a cell adhesion molecule expressed on various cell types, including activated T cells, dendritic cells, and certain tumor cells. The MEM-166 antibody can be used for the detection and analysis of CD166-expressing cells in flow cytometry and other immunoassays.

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3 protocols using mem 166

1

Immunophenotyping of CD177-Expressing Cells

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Cells were suspended in PBS with 2% FBS and incubated with CD177-FITC conjugated antibody (MEM-166; Biolegend) for 30 min. Cells were washed and evaluated by flow cytometry using a BD FACSCanto II (BD Biosciences, San Jose, CA) and analyzed using FlowJo software (FlowJo LLC, Ashland, OR).
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2

Neutrophil Characterization by Flow Cytometry

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Peripheral blood was collected in EDTA vial, and 200 μl of blood was aliquoted for antibody staining. Briefly, 1 ml of RBC lysis buffer was added to 200 μl of whole blood, and mixed properly. A 10× red blood cell lysis buffer was prepared in-house using NH4Cl (0.155 M), KHCO3 (0.01 M) and EDTA (0.1 mM). The tubes were incubated for 10 min at 4 °C, and centrifuged at 300×g for 10 min. The cell pellets were washed twice with 1× PBS and suspended in 300 μl of PBS to obtain a single cell suspension. For cell surface staining, 100 μl of cell suspension was used, and the antibodies CD16 (1:100) (clone 3G8, APC conjugated, Cat No 302011, Biolegend, USA), CD66b (1:100) (clone G10F5, FITC conjugated, Cat No 305103, Biolegend, USA) and CD177 (1.5:100) (clone MEM-166, APC/Cyanine 7 conjugated, Cat No 315809, Biolegend, USA) was used. Unstained controls, stained samples, and fluorescence minus one controls were acquired on BD-LSR Fortessa flow cytometry machine. Using the FACS DIVA software, PMN gating was done based on FSC-A v/s SSC-A plot. Enriched and activated neutrophils were gated based on CD16+ CD66b+ (double positive) in a quadrant plot. CD177+ neutrophils were gated within these double positive cells.
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3

Immunophenotyping of CD177-Expressing Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were suspended in PBS with 2% FBS and incubated with CD177-FITC conjugated antibody (MEM-166; Biolegend) for 30 min. Cells were washed and evaluated by flow cytometry using a BD FACSCanto II (BD Biosciences, San Jose, CA) and analyzed using FlowJo software (FlowJo LLC, Ashland, OR).
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