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 required that you determine the electrophoretic pattern of your protein by 1D and 2D gel electrophoresis.

We offer to separate the proteins of your drug sample. 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 analysis in combination with many of our other services. Hence, you may wish to have a look at:

If you need more information before ordering, please send us a request.

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 10-20 work days

More information:

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

Read blog post Gel blotting protocol

Order the service here

You can easily order online. If you would like guidance or a quote please contact us.

Order online

This product is available to order online. Enter your sample details and pay online or by PO/invoice

Order online

Contact us by email

Not sure wich to choose? Contact us before you order.

Open contact form

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:

Protein separation by 1D SDS-PAGE is performed using the NuPAGE pre-cast gel system from Thermo Scientific according to the manufacturer’s manual. The protein sample (5 – 20 ug) is dissolved in LDS sample buffer with DTT and heated (70 oC, 10 minutes), loaded onto the NuPAGE 12% Bis-Tris gel and separated with a MOPS/MES running buffer at 200V for 40-50 minutes. The gel is then stained 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:

2D-SDS PAGE is performed 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. The sample is purified/desalted using 2-D Clean-Up Kit (GE Healthcare). The purified sample is mixed with Destreak Rehydration solution with IPG-buffer prior isoelectric focusing. SeeBlue Plus2 Pre-Stained Standard (Invitrogen) is used as marker.

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

Finally, the 2D gels are stained by Coomassie brilliant Blue R250, or by Silver staining (Alphalyse procedure).

difference between 1d and 2d gel electrophoresis

Short 1D page, Long 1D page and 2D page

Sample preparation

Sample requirements

The samples delivered for gel electrophoresis should be delivered in-solution or as a solid material.

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
Sign up for the newsletter
Subscribe to get 3 tips that will instantly improve your protein analysis results

Receive monthly emails with news, tips, case stories
and offers for protein analysis
Private, secure, spam-free
© Copyright Alphalyse 2002 - 2020. All rights reserved.