SH-SY5Y cells were differentiated as described above and treated according to experimental design, either alone or in coculture with BV2 cells. Cultures were incubated in PBS, detached using a cell scraper, and incubated with FITC-conjugated Annexin A5 (0.45 μg/ml in 0.01 M PBS, 0.1% bovine serum albumin, and 1 mM CaCl2), and in the case of cocultures, incubated with APC-conjugated rat monoclonal anti-mouse CD11b and PerCP-Cy5.5-conjugated mouse monoclonal anti-human CD200 (all BioLegend, UK) on ice in the dark for 30 min. Samples were washed and analysed by flow cytometry. Immunofluorescence was analysed for 10,000 singlet events per sample using a FACSCanto II flow cytometer (BD Biosciences, UK); data were analysed using FlowJo 8.8.1 software (TreeStar Inc., FL, USA).
Apc conjugated rat monoclonal anti mouse cd11b
Manufactured by BioLegend
Sourced in United Kingdom
APC-conjugated rat monoclonal anti-mouse CD11b is a laboratory reagent used to detect and study the CD11b protein, which is expressed on the surface of certain immune cells in mice. The APC fluorescent dye is conjugated to the anti-CD11b antibody, allowing for the identification and analysis of CD11b-positive cells using flow cytometry or other fluorescence-based techniques.
Automatically generated - may contain errors
4 protocols using apc conjugated rat monoclonal anti mouse cd11b
1
Apoptosis Quantification in SH-SY5Y Cells
2
Immunophenotyping of Microglia and Neurons
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BV2 or SH-SY5Y cells alone or in coculture were labelled with APC-conjugated rat monoclonal anti-mouse CD11b, PE-Cy7-conjugated rat monoclonal anti-mouse CD40, or PerCP-Cy5.5-conjugated mouse monoclonal anti-human CD200 (all from BioLegend, UK) for analysis by flow cytometry. Immunofluorescence was analysed for 10,000 singlet events per sample using a FACSCanto II flow cytometer (BD Biosciences, UK); data were analysed using FlowJo 8.8.1 software (TreeStar Inc., FL, USA).
3
Apoptosis Quantification in SH-SY5Y-BV2 Co-culture
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SH-SY5Y cells were differentiated as described above and treated according to experimental design, either alone or in co-culture with BV2 cells. cultures were in PBS, detached using a cell scraper and incubated with FITC-conjugated annexin A5 (0.45 µg/ml in 0.01 M PBS, 0.1% bovine serum albumin, 1 mM CaCl2), and in the case of co-cultures, APC-conjugated rat monoclonal anti-mouse CD11b and PerCP-Cy5.5-conjugated mouse monoclonal anti-human CD200 (all Biolegend, UK) on ice in the dark for 30 min. Samples were washed and analysed by flow cytometry. Immunofluorescence was analysed for 10,000 singlet events per sample using a BD FACSCanto II (BD Biosciences, UK) flow cytometer; data were analysed using FlowJo 8.8.1 software (Treestar Inc., CA, USA).
4
Flow Cytometric Characterization of Microglial Cells
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BV2 or SH-SY5Y cells alone or in co-culture were labelled with APC-conjugated rat monoclonal anti-mouse CD11b, PE-Cy7-conjugated rat monoclonal anti-mouse CD40 (Biolegend, UK) or PerCP-Cy5.5-conjugated mouse monoclonal anti-human CD200 (all Biolegend, UK) for analysis by flow cytometry. Immunofluorescence was analysed for 10,000 singlet events per sample using a BD FACSCanto II (BD Biosciences, UK) flow cytometer; data were analysed using FlowJo 8.8.1 software (Treestar Inc., CA, USA).
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