Protein separation

Separation of proteins from sample mixture

HPLC and SDS PAGE are two of the most common protein separation techniques. They are thus used to distinguish proteins and peptides by their physiochemical properties.

Alphalyse can help you with both protein separation techniques. For instance, we can set up any customer method within our ...

HPLC and SDS PAGE are two of the most common protein separation techniques. They are thus used to distinguish proteins and peptides by their physiochemical properties.

Alphalyse can help you with both protein separation techniques. For instance, we can set up any customer method within our lab and optimize the HPLC methods for specific purposes.

Protein separation can stand alone and provide you with information about specific proteins. However, you can also combine with other analyses to purify the sample beforehand. For instance when a HPLC system is coupled to a mass spectrometer.

When to use UV-HPLC analysis:

It can also be helpful to combine the HPLC analysis with mass spectrometry. Then you can to obtain additional information about chromatographic peaks and stained spots on gels. Our protein separation services rely on more than 15 years of experience working with these analyses.

Separation by SDS PAGE gel bands:

Protein separation in SDS gel bands is often a vital part of analyses such as protein ID by mass spectrometry or N-terminal sequencing by Edman degradation. If you prefer staining and cutting gels for protein identification yourself, we recommend that you check our protocols. Please send a request for combinations of protein separation and protein analysis by mass spectrometry or Edman sequencing.

You can read more about the protein separation services below.

You may also want to combine with our other analyses: N-terminal Edman sequencing, protein identification, peptide identification, process-related impurity analysis or post-translational modifications.

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    How much protein should I load onto a gel for protein ID?

    The protein load on each gel should be around 50-300 microgram for complex samples with many proteins, and 5-50 microgram for pure protein samples.

    Should I use 1D or 2D PAGE for protein separation?

    2D PAGE is optimal to separate and visualize many proteins from complex proteomics samples. However, for large and hydrophobic proteins it is better to use 1D SDS PAGE.

    How can I increase protein concentration before loading onto 1D SDS PAGE?

    You can either use TCA precipitation, ice cold ethanol/acetone precipitation, or classical acetone precipitation.

    How do I cut the protein band/spot from an SDS PAGE gel?

    Cut the gel on a clean glass plate, while wearing gloves. Use a new clean scalpel blade. Then place the gel piece in a clean tube.

    Who is this for?

    We primarily help customers within the pharmaceutical industry, and biotechs working on drug development, e.g.

    Project managers

    Working with process development and characterization projects.

    Analytical experts

    Protein scientists that work with ELISA, masss spectrometry, or CMC.

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