Single-cell suspensions of pools of BM cells were prepared from uninfected or from 70 days M. avium 25291 infected WT or IFNγKO mice. BM progenitor cells were purified from each suspension using the Lineage Cell Depletion Mouse Kit microbeads (Miltenyi Biotec). Magnetic separation was performed with an autoMACS separator (Miltenyi Biotec). After purification, viable cells were counted by trypan blue exclusion using a hemocytometer; purity was confirmed by flow cytometry stain using FITC-conjugated anti-lineage markers and PE-conjugated anti-cKit. Cells (1-1.5 x 106) were transferred i.v. to RAGKO or iNOS.RAG.2KO mice treated with Busulfan (0.6 mg/mouse) administered i.p. 24 h before. Recipient mice received prophylactic antibiotic treatment ad libitum [2,5% of Bactrim (40 mg trimethoprim and 200 mg sulfametoxazol) in drinking water] for 5 days (treatment finished 2 days before BM transfer). Mice were euthanized 4 weeks after cell transfer and the recipient thymus and spleen were analyzed by flow cytometry.
Lineage cell depletion mouse kit microbeads
Manufactured by Miltenyi Biotec
The Lineage Cell Depletion Mouse Kit microbeads are designed for the depletion of unwanted cell populations from mouse cell suspensions. The microbeads target specific cell surface antigens, allowing for the selective removal of those cells from the sample.
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Lab products found in correlation
2 protocols using lineage cell depletion mouse kit microbeads
1
Reconstitution of Adaptive Immunity in Mice
2
Adoptive Transfer of Bone Marrow Progenitors
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Single-cell suspensions of pools of BM cells were prepared from non-infected or from 70 days M. avium 25291 infected WT or IFNgKO mice. BM progenitor cells were purified from each suspension using the Lineage Cell Depletion Mouse Kit microbeads (Miltenyi Biotec).
Magnetic separation was performed with an autoMACS separator (Miltenyi Biotec). After purification, viable cells were counted by trypan blue exclusion using a hemocytometer; purity was confirmed by flow cytometry stain using FITC-conjugated anti-lineage markers and PEconjugated anti-cKit. Cells (1-1.5 x 10 6 ) were transferred i.v. to each RAGKO or iNOS.RAG.2KO mouse treated with Busulfan (0.6mg/mouse) i.p. 24 hours before. Recipient mice received prophylactic antibiotic treatment ad libitum [2,5% of Bactrim (40 mg trimethoprim + 200 mg sulfametoxazol) in drinking water] for 5 days (treatment finished 2 days before BM transference). Mice were euthanized 4 weeks after cell transfer and the recipient thymus and spleen were analyzed by flow cytometry.
Magnetic separation was performed with an autoMACS separator (Miltenyi Biotec). After purification, viable cells were counted by trypan blue exclusion using a hemocytometer; purity was confirmed by flow cytometry stain using FITC-conjugated anti-lineage markers and PEconjugated anti-cKit. Cells (1-1.5 x 10 6 ) were transferred i.v. to each RAGKO or iNOS.RAG.2KO mouse treated with Busulfan (0.6mg/mouse) i.p. 24 hours before. Recipient mice received prophylactic antibiotic treatment ad libitum [2,5% of Bactrim (40 mg trimethoprim + 200 mg sulfametoxazol) in drinking water] for 5 days (treatment finished 2 days before BM transference). Mice were euthanized 4 weeks after cell transfer and the recipient thymus and spleen were analyzed by flow cytometry.
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