1D and 2D SDS PAGE – gel electrophoresis

  • Get the electrophoretic pattern of your protein sample
  • Combine analysis with other protein services
  • Purify and prepare a complex sample for mass spectrometry

Do you need one- or two-dimensional gel electrophoresis?

According to the ICH Q6B (Test Procedures and Acceptance Criteria for Biotechnological/Biological Products) i...

Do you need one- or two-dimensional gel electrophoresis?

According to the ICH Q6B (Test Procedures and Acceptance Criteria for Biotechnological/Biological Products) it is a requirement that you determine the electrophoretic pattern of your protein by 1D and 2D gel electrophoresis.

If you need help, we can separate the proteins of your drug sample for you. This is done by either 1D PAGE or 2D PAGE (polyacrylamide gel electrophoresis). We use Coomassie, silver or fluorescent staining for visualization of all proteins in the sample.

Our gel electrophoresis service includes:

You can order our 1D PAGE and 2D gel electrophoresis in combination with one of our many other analysis services. Hence, you may wish to have a look at:

If you need more information before ordering, please send us a request using the form below.

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The process:

  • 1 You provide us with in-solution or solid samples
  • 2 We perform 1D- or 2D gel electrophoresis and stain the proteins with a suitable method
  • 3 Optional: We analyze the separated proteins
  • 4 You receive results within 10-20 work days

More information:

1D SDS PAGE versus 2D PAGE. What are the recommendations?

Read blog post Gel blotting protocol - PVDF membrane

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Knowledge center

More information

We would like to help you as much as possible with your project and therefore provide several kinds of customer support:

Technical details

Introduction to 1D and 2D gel electrophoresis

1D gels:

We perform protein separation by 1D SDS-PAGE with the NuPAGE pre-cast 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 onto the NuPAGE 12% Bis-Tris gel and separate with a MOPS/MES running buffer at 200V for 40-50 minutes. Finally, we stain the gel by Coomassie brilliant Blue R250, or by Silver staining (Alphalyse procedure).

loading of 1d sds page for gel electrophoresis

1D page separation with PVDF blotting for N-terminal sequencing of different DS batches

 

2D gels:

We perform 2D-SDS PAGE using IPG drystrips 7 cm, pH 3-10 for the first dimension isoelectric focusing, and NuPAGE pre-cast gel system from Invitrogen for the second dimension of Mw separation. First, the sample is purified/desalted using 2-D Clean-Up Kit (GE Healthcare). Then we mix the purified sample with Destreak Rehydration solution with IPG-buffer prior isoelectric focusing. Also, we use SeeBlue Plus2 Pre-Stained Standard (Invitrogen) as marker.

  1. The isoelectric focusing runs until approximately 8000 Vhrs.
  2. The second dimension uses NuPAGE precast 4-12% Bis-Tris zoom gels, and proteins separate with a MOPS/MES running buffer at 200V for 40-50 minutes.

Finally, we stain the 2D gels by either Coomassie brilliant Blue R250, or by Silver staining (Alphalyse procedure).

difference between 1d and 2d gel electrophoresis

Examples of short 1D page, Long 1D page and a 2D page

Sample preparation

Sample requirements

It is important that the samples delivered for gel electrophoresis are 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.

Meet the experts

For this type of analysis our experts include:

Do you need help?
Thanh Ha Nguyen

MSc in Biotechnology

Read more
Do you need help?
Martin Villadsen

MSc in Engineering within Medical Biotechnology

Read more
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