Bone marrow cells flushed from long bones were syringed five times through a 23-G needle and filtered through a 100μm nylon cell strainer. Red blood cells were lysed with 0.15 M NH4Cl. Cells were resuspended in PBS and filtered through a 70μm strainer. Samples were incubated with Fixable Viability Dye eFluor780 (eBioscience, San Diego, CA) and Fc receptors were blocked with anti-mouse CD16/CD32 (Biolegend, San Diego, CA) at a dilution of 1:100. Cells were then incubated with anti-mouse CD115 APC, anti-mouse CD11b PERCP-Cy5.5, anti-mouse B220 Pe-Cy7, anti-mouse CD11c PE, and anti-mouse Gr-1 Brilliant Violet 421 (BioLegend) at a dilution of 1:100 in PBS/2% FBS. Samples were acquired on a FACSCanto II or LSR II SORP (BD Biosciences, San Jose, CA). Analysis was performed with FlowJo software (Tree Star, Ashland, OR).
Anti mouse b220 pe cy7
Manufactured by BioLegend
Sourced in United States
Anti-mouse B220 PE-Cy7 is a fluorochrome-conjugated monoclonal antibody that binds to the B220 (CD45R) antigen expressed on mouse B cells. It can be used for the identification and enumeration of B cells in flow cytometry applications.
Automatically generated - may contain errors
Lab products found in correlation
Lsrii sorp, by BD (1 mentions)
Anti mouse cd16 cd32, by BioLegend (1 mentions)
Anti mouse cd115 apc, by BioLegend (1 mentions)
Pe anti mouse cd11c, by BioLegend (1 mentions)
Anti mouse cd11b percp cy5, by BioLegend (1 mentions)
Fixable viability dye efluor 780, by Thermo Fisher Scientific (1 mentions)
Facscanto 2, by BD (1 mentions)
Anti cd16 cd32, by BD (1 mentions)
Igm fitc, by BioLegend (1 mentions)
Igd apc, by BioLegend (1 mentions)
Cd38 pe, by BioLegend (1 mentions)
Facs attune, by Thermo Fisher Scientific (1 mentions)
2 protocols using anti mouse b220 pe cy7
1
Isolation and Characterization of Murine Myeloid Cells
2
Profiling Memory B Cells from Vaccination
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To quantify and profile memory B cells from vaccination, single‐cell suspensions were prepared from inguinal lymph nodes, as previously described.32 Cells were stained with Zombie live/dead (Biolegend, San Diego, CA, USA), FcR‐blocked (anti‐CD16/CD32, BD Bioscience, Franklin Lakes, NJ, USA) and stained with a cocktail containing anti‐mouse B220‐PECy7, CD38‐PE, IgD‐APC, CD95 (Fas)‐BV605, IgM‐FITC, IgG‐APCCy7 (all Biolegend), for 30 min on ice in FACS buffer (PBS, 1% FBS, 0.5% NaN3). Cells were finally fixed with 100 µL of 4% PFA for 20 min on ice. B memory cells were defined as B220+ CD38+/− Faslow IgD‐ IgM+ or IgG+. Samples were acquired by flow cytometry on a FACS Attune (Invitrogen) and analysed with FlowJo software (BD). Representative FACS plots are shown in Supplementary figure 2 a.
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