Bone Marrow Immune Cell Profiling

Bone marrow cells were flushed and collected from femurs (n = 8 to 10 per group). An amount of 1 × 10⁶ cells was plated per well in a 96-well plate. The staining was performed as described by Collins et al. (43 (link)). Briefly, cells were blocked with Fc block (Becton Dickinson Pharmingen, Franklin Lakes, NJ) for 15 min and then stained with antibodies to CD3 (catalog number 56-0032; eBioscience, San Diego, CA), CD4 (catalog number 11-0041; eBioscience, San Diego, CA), CD8a (catalog number 35-0081; eBioscience, San Diego, CA), GR1/Lys-6G (catalog number 53-5931; eBioscience, San Diego, CA), and CD11b (catalog number 12-0113; eBioscience, San Diego, CA) for 30 min. Then, cells were fixed and data were acquired using the Becton Dickinson LSRII flow cytometer. Data analysis was performed using the FlowJo software package (FlowJo, LLC).

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Quach D., Collins F., Parameswaran N., McCabe L, & Britton R.A. (2018). Microbiota Reconstitution Does Not Cause Bone Loss in Germ-Free Mice. mSphere, 3(1), e00545-17.

Publication 2018

Fc block CD11b LSRII flow cytometer FlowJo software package

Antibodies Bone marrow cells Cd11b Cells Femurs

Corresponding Organization :

Other organizations : Michigan State University, Baylor College of Medicine

Top 5 similar protocols

Variable analysis

independent variables

N = 8 to 10 per group

dependent variables

Cell populations measured by flow cytometry: CD3, CD4, CD8a, GR1/Lys-6G, CD11b

control variables

1 × 10^6 cells plated per well in a 96-well plate
Cells were blocked with Fc block for 15 min before staining
Cells were stained with antibodies for 30 min
Cells were fixed before data acquisition

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