I have to isolate different T-cell and B-Cells population from mice tumors. Can anyone please suggest me markers for flow cytometry and immunohistochemistry.
To add to Stephane's answer: CD62L is exceedingly heat labile and will easily detach from T cells. Thus, be very careful to ensure temperature stability (4 degrees) during the cell manipulation process. In general, it is my understanding that the following scheme is most common:
CD62L+CD44lo or just CD44 lo in the event that CD62L is cleaved (naive) CD62L+CD44+i (central memory) CD62L-CD44+ (effector memory). CCR7 is a very helpful addition.
For T cell:
CD3, CD4, CD8 if you want to study regulatory cells then:
CD3+ CD4+ CD25+ AND FOXP3+
For B cells:
B220, CD19, CD20
If you want to study other types of B or T cells there are more specific markers.
For Immunohistochemistry same markers are used.
Hope this can help
http://www.abcam.com/primary-antibodies/b-cells-basic-immunophenotyping
http://www.abcam.com/primary-antibodies/t-cells-basic-immunophenotyping
I would recommend the following combination:
T cells: CD3 + CD5 or CD3 + CD8,
B cells: CD19+B220
If you are searching for specific cell types you can find a detailed information in the links below:
https://www.bdbiosciences.com/documents/cd_marker_handbook.pdf
https://www.bio-rad-antibodies.com/b-cell-function-activation-lineage-marker-antibody.html
https://static.bio-rad-antibodies.com/2016/biomarker-resources/biomarker-expression-patterns-in-mouse-immune-cells.pdf
https://www.bio-rad-antibodies.com/static/2015/mouse-poster/mouse-immune-cell-lineage-antigen-expression-poster.pdf
I don’t think CD 5 can be a useful marker for identifying T cell population in Mice.
CD 5 is usually used for identifying B-1a cells.
Hope Dr. Dimitar Parvanov will agree on this.
To Prashant: T cells are indeed CD5hi while B1a cells are CD5lo
Markers of B cells in periphery (CD19+B220+ for all except Plasma cells B220loCD19lo)
Naive B cells: CD21+CD23+IgD+CD38hi
MZ B cells: CD21+CD23-IgMhi
GC B cells:GL7+CD38loCXCR4+ (centroblasts) CXCR4- (centrocytes)
Plasma cells: CD138+ IgM+ (unswitched) IgG+ (switched) CD93+ (Plasmablasts and long term PC)
Markers of T cells:
Helper T: CD4+ CD3+ TCRab+
cytotoxic T: CD8+ CD3+ TCRab+
CD62L+CD44lo (naive) CD62L-CD44lo (effector) CD62L-CD44+ (memory)
Tregs CD4+CD25+
Good luck!
Dear Dr. Stephane J C Mancini,
Thanking you very much for the information.
To add to Stephane's answer: CD62L is exceedingly heat labile and will easily detach from T cells. Thus, be very careful to ensure temperature stability (4 degrees) during the cell manipulation process. In general, it is my understanding that the following scheme is most common:
CD62L+CD44lo or just CD44 lo in the event that CD62L is cleaved (naive) CD62L+CD44+i (central memory) CD62L-CD44+ (effector memory). CCR7 is a very helpful addition.
