Cells were stained in PBS supplemented with 0.5% BSA and 2mM EDTA. Ghost Dye Red 780 (Tonbo biosciences, #13–0865) was used to discriminate viable from non-viable cells, then staining with Anti-Mouse CD16/CD32 (Clone 2.4G2, BD Pharmingen #553142) to inhibit non-specific staining, following manufacturer’s instructions. Fluorochrome-conjugated mAbs specific to mouse CD169-BV605 and Alexa 647 (clone 3D6.112), F4/80-PerCP-Cy5.5 (clone BM8), Ter119-APC (clone TER-119), CD71-FITC (clone RI7217) and CD11b- BV650 (clone M1/70), all purchased from BioLegend. Cell suspensions were fixed and acquired on Becton-Dickinson LSRFortessa and analyzed using FlowJo software (Tree Star).
Cd11b bv650 clone m1 70
Manufactured by BioLegend
CD11b-BV650 (clone M1/70) is a flow cytometry reagent used to detect the expression of the CD11b cell surface antigen. CD11b is a member of the integrin family and is expressed on the surface of various leukocyte populations, including monocytes, macrophages, and neutrophils. The BV650 fluorochrome is conjugated to the CD11b antibody, allowing for the detection of CD11b-positive cells by flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Anti mouse cd16 cd32, by BD (1 mentions)
Lsr fortessa, by Tree Star (1 mentions)
Ghost dye red 780, by Cytek Biosciences (1 mentions)
Fluorochrome conjugated mabs specific, by BioLegend (1 mentions)
Ly6g apc cy7, by BioLegend (1 mentions)
Gentlemacs instrument, by Miltenyi Biotec (1 mentions)
Cd11c bv605, by BioLegend (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Siglec f bv421, by BD (1 mentions)
Lsrii flow cytometer, by BD (1 mentions)
2 protocols using cd11b bv650 clone m1 70
1
Staining for Splenic Myeloid Cells
2
Comprehensive Lung Cell Isolation
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Whole lungs were removed at 24 hours post-infection and dissociated into a single cell suspension per manufacturer’s protocol (Miltenyi 130-095-927). Briefly, separated lung lobes were placed in a C tube (Miltenyi 130-096-334) containing 2.4 ml 1X Buffer S, 100 µl enzyme D, and 15 µl enzyme A. Samples were mechanically disrupted using a GentleMACS instrument (Miltenyi), filtered, and red blood cells were lysed by incubation in Ammonium-Chloride-Potassium (ACK) buffer (Gibco A10492-01) (16 (link)). 106 cells per sample were stained in PBS (Gibco) + 1% BSA (Quality Biological Inc K719500ML) with the following antibody cocktail: CD45-FITC (clone 30-F11, Biolegend 103108), CD11b-BV650 (clone M1/70, Biolegend 101259), CD11c-BV605 (clone N418, Biolegend 117334), F4/80-PerCP-Cy5.5 (clone BM8, Biolegend 123128), SiglecF-BV421 (clone E50-2440, BD Horizon 562681), and Ly6G-APC-Cy7 (clone 1A8, Biolegend 127624). Samples were resuspended in stabilizing fixative (BD 338036), run on an LSRII flow cytometer (BD Biosciences), and data were analyzed using FlowJo software v10.7.1 (BD Life Sciences).
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