Quantification of secreted protein in conditioned media.

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Renumbered during migration. The source used a single continuous numbering across both sections and had two steps numbered 14. The sequence is unchanged.

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Materials

• Conditioned media stored at -80°C
• AcroPrep Advance 0.2 μm Supor 96-well filter plate (VWR 97052-124)
• AcroPrep Advance 3K Omega 96-well filter plate (VWR 89135-696)
• 96-well deep well plate (VWR 76210-518)
• Troughs
• PBS
• NanoOrange Assay Kit (Fisher N6666)
• 96-well PCR plate
• Tin foil

Equipment

• Eppendorf benchtop centrifuge
• SpectraMax I3x multi-mode plate reader
• SpeedVac (optional)
• 12-channel pipettes
• Thermocycler

Reference material: ThermoFisher NanoOrange manual and Molecular Devices application note.

Protein purification