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Isoelectric Focusing: A Simple Way to Enhance Your Protein Separation
Manage episode 435212165 series 3391757
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#112 — Isoelectric focusing (IEF) is a powerful technique distinct from the more familiar SDS-PAGE, [1,2] tailored for separating proteins or peptides based on their isoelectric points (pI). [3]
This method capitalizes on the migration of charged molecules through a stable pH gradient until they reach a zone where their net charge is zero, halting their movement.
Essential to setting up an IEF experiment are the IPG strips, equipped with a pH-responsive gel, and the sample, often mixed with carrier ampholytes for efficient migration.
IEF's utility spans various applications, from enhancing 2D-PAGE protein separations to analyzing post-translational modifications and preparing samples for mass spectrometry. [4]
Discover more about how this technique can advance your research and streamline your experimental workflows.
Resources:
1. How SDS-PAGE Works. Available at: https://bitesizebio.com/580/how-sds-page-works/
2. Isoelectric Focusing: A Simple Way to Enhance Your Protein Separation. Available at: https://bitesizebio.com/26811/isoelectric-focusing-separation-proteins-peptides/
3. The Definitive Guide to pH, pKa, and pI. Available at: https://bitesizebio.com/9425/what-is-ph-pka-pi/
4. Detecting Post-Translational Modifications. Available at: https://bitesizebio.com/49322/detecting-post-translational-modifications/
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