Fc receptors were blocked with anti-mouse CD16/CD32 for 10 min on ice. Cells were then incubated for 30 min on ice with CD11b PE, CD45 FITC, and Ly-6C APC (Abcam) and subsequently washed with Medium A without Phenol Red and sorted using a BD FACSAria III cell sorter (Becton Dickinson, Franklin Lakes, NJ, USA). Microglia were defined as CD11bpos CD45int Ly-6Cneg [32 (link)] and determined by flow cytometry with a mean purity of 99%. Viability was tested using trypan blue exclusion dye. Finally, cells were suspended at 1 × 105 cells/ml in DMEM supplemented with 10% FBS.
Cd11b pe
Manufactured by Abcam
Sourced in United States, United Kingdom
CD11b PE is a fluorescently-labeled antibody that binds to the CD11b antigen. CD11b is a cell surface integrin protein expressed on various immune cells, including monocytes, macrophages, and neutrophils. This product can be used for the identification and analysis of CD11b-positive cells in flow cytometry applications.
Automatically generated - may contain errors
2 protocols using cd11b pe
1
Isolation and Purification of Microglia
2
Flow Cytometry Analysis of Cultured Cells
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The third-generation cultured cells were digested with 0.25% trypsin for 5 minutes. After centrifugation at 1,000 rpm for 10 minutes, the cells were washed twice with PBS, and then the cells were resuspended with PBS to prepare cell suspensions. The cell concentration was adjusted to 2×106/mL. The cells were divided into flow centrifuge tubes at 500 µL per tube, and CD11B-PE (Abcam Cambridge, UK), CD29-PE (Abcam, UK), CD-34-FITC (Abcam, UK), CD45-PE (Abcam, UK), CD90-PE (Abcam, UK), CD105 PE (Abcam, UK), and the isotype control (Abcam, UK) were added. The amount of each added antibody was 10 µL. After incubation at 4 °C for 30 minutes, cells were washed twice with PBS, then 4% paraformaldehyde (Guangzhou Jibeisi Biological Technology Co., Ltd.) at 200 µL was added to each tube to resuspend the cells. A filter membrane was used to filter cell masses to prevent blockage of the flow cytometer. Cells were then fed into the flow cytometer for identification.
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