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Set up meetingProtein characterization for optimization of manufacturing processes.
Explore customer caseHigh-throughput characterization analysis of antibodies for fast quality assessment
Explore customer caseWe would like to help you as much as possible with your project and therefore provide several kinds of customer support:
We perform protein separation by 1D SDS-PAGE with the NuPAGE precast gel system from Thermo Scientific and according to the manufacturer’s manual. First, we dissolve the protein sample (5 – 20 ug) in LDS sample buffer with DTT and heat (70 oC, 10 minutes). Then we load it onto the NuPAGE 12% Bis-Tris gel and separate it with a MOPS/MES running buffer at 200V for 40-50 minutes. Finally, we stain the gel with Coomassie Brilliant Blue R250 or Silver staining (Alphalyse procedure).
1D page separation with PVDF blotting for N-terminal sequencing of different DS batches
First, the sample is purified/desalted using a 2-D Clean-Up Kit (GE Healthcare). Then we mix the purified sample with Destreak Rehydration solution with IPG-buffer before isoelectric focusing. We perform 2D-SDS PAGE using IPG dry strips 7 cm, pH 3-10 for the first dimension isoelectric focusing, and NuPAGE precast gel system from Invitrogen for the second dimension of Mw separation. Also, we use SeeBlue Plus2 Pre-Stained Standard (Invitrogen) as a marker.
Finally, we stain the 2D gels with either Coomassie brilliant Blue R250 or Silver staining (Alphalyse procedure).
Examples of a short 1D page, a Long 1D page, and a 2D page
The samples delivered for gel electrophoresis must be in-solution or solid material.
Also, note that for 1D gels, we will typically load between 2 and 30 µg, and for 2D gels, between 30 and 300 µg.
