All naive RAG1−/− mice were i.v. reconstituted with 5×106 CD4+ T cells. CD4+ T cells are isolated by first B220+ depletion, followed by anti-mouse L3T4 magnetic bead selection (Miltenyi Biotec) from the spleens of WT mice. Indicated mice were i.v. reconstituted with 10 × 106 naive B2 cells. B2 cells are isolated by depletion using either anti-mouse CD43 Miltenyi microbeads (Miltenyi Biotec) or biotinylated anti-mouse CD43 (AB_493384) followed by anti-biotin microbeads (Miltenyi Biotec) from the spleens of WT mice. Indicated mice were i.p. reconstituted with 2–4 × 105 B1 cells. B1 cells are isolated from the peritoneal and pleural cavities of WT mice (Yenson and Baumgarth, 2014 (link)). Briefly, cells were Fc blocked on ice for 10 min (2.4G2) (Unkeless, 1979 (link)) followed by the following biotinylated antibodies for 30 min on ice: anti-mouse CD23 (B3B4) (AB_312829), anti- mouse CD49b (DX5) (AB_313035), anti-mouse F4/80 (BM8) (AB_893499), anti-mouse CD90.2 (30-H12) (AB_313175), and anti-mouse GR-1 (RB6-8C5) (AB_313368). After washing, anti-biotin microbeads (Miltenyi Biotec) were added, and magnetic bead depletion was performed for B1 enrichment. Mice were not used in experiments until 1 week post reconstitution.
Anti mouse l3t4 magnetic bead selection
Manufactured by Miltenyi Biotec
The Anti-mouse L3T4 magnetic bead selection is a lab equipment product used for the isolation and enrichment of CD4+ T cells from mouse samples. It utilizes magnetic beads coated with antibodies specific to the L3T4 (CD4) surface marker to capture and separate the target cell population.
Automatically generated - may contain errors
2 protocols using anti mouse l3t4 magnetic bead selection
1
Adoptive Transfer of T and B Cells in RAG1-/- Mice
2
Adoptive Transfer of T and B Cells in RAG1-/- Mice
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All naive RAG1−/− mice were i.v. reconstituted with 5×106 CD4+ T cells. CD4+ T cells are isolated by first B220+ depletion, followed by anti-mouse L3T4 magnetic bead selection (Miltenyi Biotec) from the spleens of WT mice. Indicated mice were i.v. reconstituted with 10 × 106 naive B2 cells. B2 cells are isolated by depletion using either anti-mouse CD43 Miltenyi microbeads (Miltenyi Biotec) or biotinylated anti-mouse CD43 (AB_493384) followed by anti-biotin microbeads (Miltenyi Biotec) from the spleens of WT mice. Indicated mice were i.p. reconstituted with 2–4 × 105 B1 cells. B1 cells are isolated from the peritoneal and pleural cavities of WT mice (Yenson and Baumgarth, 2014 (link)). Briefly, cells were Fc blocked on ice for 10 min (2.4G2) (Unkeless, 1979 (link)) followed by the following biotinylated antibodies for 30 min on ice: anti-mouse CD23 (B3B4) (AB_312829), anti- mouse CD49b (DX5) (AB_313035), anti-mouse F4/80 (BM8) (AB_893499), anti-mouse CD90.2 (30-H12) (AB_313175), and anti-mouse GR-1 (RB6-8C5) (AB_313368). After washing, anti-biotin microbeads (Miltenyi Biotec) were added, and magnetic bead depletion was performed for B1 enrichment. Mice were not used in experiments until 1 week post reconstitution.
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