1000 sort-purified LT-HSCs from M2/HSB/Tn mice were cultured together with 10,000 MS-5 stromal cells in round-bottom 96-well plates together with SCF (100 ng/ml), TPO (100 ng/ml), Flt3L (50 ng/ml), IL7 (20 ng/ml), IL3 (10 ng/ml), IL11 (50 ng/ml), and GM-CSF (20 ng/ml) in αMEM with 1% Penicillin/Streptomycin and 10% FCS (Thermo Fisher) for two weeks, changing the media 24h after sort and then every 48h (Becton Dickinson). Myeloid and lymphoid HSC progeny was FACS sorted after labelling with Gr-1, Mac-1, CD19 and B220 antibodies (eBiosciences). All growth factors and cytokines were mouse recombinant and purchased from Peprotech.
B220 antibodies
Manufactured by Thermo Fisher Scientific
The B220 antibodies are a type of immune cell marker used in laboratory research. They specifically recognize and bind to the CD45R protein expressed on the surface of B lymphocytes, a type of white blood cell. These antibodies are commonly used in flow cytometry and other immunological techniques to identify and quantify B cells within a sample.
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2 protocols using b220 antibodies
1
Expansion of Murine Hematopoietic Stem Cells
2
Expansion of Murine Hematopoietic Stem Cells
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1000 sort-purified LT-HSCs from M2/HSB/Tn mice were cultured together with 10,000 MS-5 stromal cells in round-bottom 96-well plates together with SCF (100 ng/ml), TPO (100 ng/ml), Flt3L (50 ng/ml), IL7 (20 ng/ml), IL3 (10 ng/ml), IL11 (50 ng/ml), and GM-CSF (20 ng/ml) in αMEM with 1% Penicillin/Streptomycin and 10% FCS (Thermo Fisher) for two weeks, changing the media 24h after sort and then every 48h (Becton Dickinson). Myeloid and lymphoid HSC progeny was FACS sorted after labelling with Gr-1, Mac-1, CD19 and B220 antibodies (eBiosciences). All growth factors and cytokines were mouse recombinant and purchased from Peprotech.
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