Human/Mouse/Rat/Canine ALCAM/CD166 Antibody

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In order to confirm the identity of harvested cells as mesenchymal progenitors, immunohistochemical staining (IHC) for mesenchymal stem cell (MSC) markers CD-105 (Endoglin), CD-166 (ALCAM), and CD-133 was performed. Briefly, harvested cells were seeded onto 12-well cell culture plates and allowed to reach 100% confluency. They were fixed with 10% formalin and washed twice with PBS. Staining was performed using anti-CD105 mouse monoclonal antibody, goat anti-CD133 polyclonal antibody, and rabbit anti-CD166 polyclonal antibody (1:100; Santa Cruz Biotechnology, Santa Cruz, CA, USA). Respective biotin-conjugated secondary antibodies were applied at a dilution of 1:600 (Santa Cruz Biotechnology) and incubation with horseradish peroxidase-strepavidin was performed at 1:300 dilution (Santa Cruz Biotechnology). The presence of the proteins of interest was visualized by DAB staining. Minus primary antibodies were used as negative controls.

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