Bone marrow was isolated from tibiae and femora, six weeks following ovariectomy. Following lysis of red blood cells using RBC lysis buffer (Sigma, St. Louis, MO, USA), single cell suspensions were prepared by passing the cells through 100μm Nytex mesh. Cells were stained with CD11b conjugated to FITC and F4/80 conjugated to PE (Invitrogen, Camarillo, CA) or appropriate isotype controls (BD Pharmingen, San Jose, CA) using staining medium (1X Hank’s balanced salt solution containing 10mM HEPES and 2% new born calf serum). Dead cells were identified and excluded by using propidium iodide staining. FACS analysis was performed using BD FACS Calibur (Franklin Lakes, NJ).
Cd11b conjugated to fitc
Manufactured by Thermo Fisher Scientific
CD11b conjugated to FITC is a lab equipment product that can be used for the detection and analysis of CD11b-expressing cells. CD11b is a cell surface marker that is expressed on various immune cells, including monocytes, macrophages, and neutrophils. The FITC (Fluorescein Isothiocyanate) fluorescent dye is conjugated to the CD11b antibody, allowing for the visualization and identification of CD11b-positive cells through flow cytometry or other fluorescence-based techniques.
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Lab products found in correlation
Appropriate isotype controls, by BD (1 mentions)
Rbc lysis buffer, by Merck Group (1 mentions)
Facscalibur, by BD (1 mentions)
Multi tissue dissociation kit 1, by Miltenyi Biotec (1 mentions)
Fortessa x20 flow cytometer, by BD (1 mentions)
Countess automated cell counter, by Thermo Fisher Scientific (1 mentions)
Macs buffer, by Miltenyi Biotec (1 mentions)
2 protocols using cd11b conjugated to fitc
1
Isolation and Characterization of Murine Bone Marrow Monocytes
2
Flow Cytometric Analysis of GBS-Infected Mouse Brains
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At 48 h post-infection with 1 × 108 CFU of GBS, mice were euthanized then perfused to replace blood with PBS. The entire mouse brain was harvested from each animal and the tissue was processed with the Multi-Tissue Dissociation kit #1 following the Adult brain dissociation protocol (Miltenyi Biotec). Cells were resuspended in MACS buffer (Miltenyi Biotec) and incubated with antibodies to Ly6C conjugated to BV421, CD45 conjugated to PE, CD11b conjugated to FITC, and Ly6G conjugated to APC (Invitrogen) at 1:200 dilution, UltraLeaf anti-mouse CD16/CD32 Fc block (Biolegend) at 1:400 dilution, and fixable viability dye conjugated to eFLuor506 (eBioscience) at 1:1000 dilution for 1 h, then fixed (eBioscience). Cells were counted using a Countess automated cell counter (Invitrogen), read on a Fortessa X-20 flow cytometer (BD Biosciences), and analyzed using FlowJo (v10) software. Gates were drawn according to fluorescence minus one (FMO) controls.
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