Frozen skin sections were prepared from 24 h cultured skin biopsies from the ‘bath’ and injection approach and fixed in acetone (VWR, Darmstadt, Germany) for 10 min at room temperature. The targeting mAbs against Langerin and DEC-205 were visualized with anti-mouse-Ig-Alexa 594 (Invitrogen) followed by blocking residual free anti-mouse binding sites with 100 μg/ml mouse gamma globulin (Jackson ImmunoResearch Laboratories, Pennsylvania, USA) for 15 min. Skin DC were identified with biotinylated anti-CD1a (clone HI149, BioLegend) and biotinylated anti-HLA-DR (clone L243, BioLegend), followed by Streptavidin-Alexa Fluor 488 (Invitrogen). Appropriate isotype controls were purchased from Biolegend. All staining steps were performed for 30 min at 37°C.
Biotinylated anti cd1a clone hi149
Manufactured by BioLegend
Biotinylated anti-CD1a (clone HI149) is a monoclonal antibody that binds to the CD1a protein. CD1a is a cell surface glycoprotein that functions in the presentation of lipid and glycolipid antigens to T cells. The biotinylation of the antibody allows for detection and binding to streptavidin or avidin-conjugated molecules.
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Lab products found in correlation
2 protocols using biotinylated anti cd1a clone hi149
1
Staining and visualization of skin DCs
2
Quantifying Skin Dendritic Cells
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Frozen skin sections were prepared from 24 h cultured skin biopsies from the ‘bath’ and injection approach and fixed in acetone (VWR, Darmstadt, Germany) for 10 min at room temperature. The targeting mAbs against Langerin and DEC-205 were visualized with anti-mouse-Ig-Alexa 594 (Invitrogen) followed by blocking residual free anti-mouse binding sites with 100 μg/ml mouse gamma globulin (Jackson ImmunoResearch Laboratories, Pennsylvania, USA) for 15 min. Skin DC were identified with biotinylated anti-CD1a (clone HI149, BioLegend) and biotinylated anti-HLA-DR (clone L243, BioLegend), followed by Streptavidin-Alexa Fluor 488 (Invitrogen). Appropriate isotype controls were purchased from Biolegend. All staining steps were performed for 30 min at 37°C.
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