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), you must determine the electrophoreti...

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

According to the ICH Q6B (Test Procedures and Acceptance Criteria for Biotechnological/Biological Products), you must 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. It 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:

  • 1D gel or 2D gel electrophoresis separation of your protein sample
  • Coomassie, silver or fluorescence staining of the proteins

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

Please send us a request using the form below to order or get more information.

MoreLess details

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 20-28 days

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We offer customized solutions, contact us to discuss your project.

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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 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).

loading of 1d sds page for gel electrophoresis

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

 

2D gels:

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.

  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 with either Coomassie brilliant Blue R250 or Silver staining (Alphalyse procedure).

difference between 1d and 2d gel electrophoresis

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

Sample preparation

Sample requirements

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.

Related services

Meet the experts

For this type of analysis our experts include:

Do you need help?
Thanh Ha Nguyen

MSc in Biotechnology

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Do you need help?
Line Koch

PhD in Health and Medical Sciences

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