Dilute 2µg of native human TPO (antibodies-online, ABIN934717, lot A17091521) in 2x sample buffer (126mM Tris-HCl pH6.8, 20% glycerol, 4% SDS, 0.02% bromophenol blue, 0.1M DTT).
Boil samples for 5min at 95°C.
Separate the sample on a denaturing SERVAGel TG PRiME 12% precast gel (Serva, 43266, V170144) in a Electrophoresis chamber (Hoefer Inc., SE250) with Laemmli buffer for 1h at 250V, 50mA/gel along side 4µl Roti-Mark Tricolor (Carl Roth, 8271, lot 326248051) molecular weight marker.
Staining of the separated proteins with InstantBlue Protein Stain (Expedeon, ISB1L, lot 170616330) according to the manufacturer’s recommendations.
Image acquisition: 600dpi scan and increasing of color saturation.
The manufacturer’s information concerning the protein’s purity and molecular weight can be confirmed.