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The MC-480 is a laboratory instrument designed for the cultivation and maintenance of hybridoma cell lines. It features a temperature-controlled incubation chamber and programmable settings to ensure optimal growth conditions for the cells.

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2 protocols using mc 480

1

Stem Cell Surface Antigen Identification

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TROMA-1, a rat monoclonal antibody against cytokeratin Endo A, a marker of primitive endoderm, and a mouse monoclonal antibody (mAb MC-480) that recognizes the stem cell-specific cell surface antigen (SSEA-1), were purchased from the Developmental Studies Hybridoma Bank (University of Iowa, IA, USA). All trans-retinoic acid (RA) was purchased from Sigma (USA) and was dissolved in ethanol (100 mM stock). Puromycin was purchased from Sigma (USA). The plasmid vector pCDNA3, Neomycin (G418) were purchased from Invitrogen (USA). Plasmid harboring non-targeting short hairpin RNA (shRNA), pLKO.1 control vector was purchased from Open Biosystems, USA.
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2

SSEA-1 Expression Analysis by Flow Cytometry

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Cells were trypsinized, washed once in PBS, and resuspended in PBS; dead cells were stained using the LIVE/DEAD Fixable Dead Cell Stain Kit (Invitrogen) according to the manufacturer’s instructions. Next, cells were fixed in 2% paraformaldehyde (PFA) for 15 min at room temperature, washed with PBS and stained with anti-SSEA-1 mouse monoclonal IgM antibody (mc-480, University of Iowa, Developmental Studies Hybridoma Bank, 1:400) at 37°C for 20 min. After a PBS wash, the cells were labeled with fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgM (Millipore, 1:200) for 15 min at 37°C. After one PBS wash, stained cells were resuspended in PBS containing 1% FBS and 3% BSA, and at least 10,000 events of healthy cells were analyzed with a CyAN Analyzer (Beckman Coulter) and FlowJo ver.10 software (Tree Star, Ashland, USA). Control cells were not treated with primary antibody.
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