Is there another method (other than SDS-PAGE) can be used for checking protein's purity?
Hello,
Besides Native/Denaturing PAGE and MS, you can also use:
- Dynamic Light Scattering (DLS) - for the homogeneity assessment,
- HPLC - for separation, identification, and quantification of compounds in a sample.
Additionally, if you suspect the presence of specific impurities, e.g., DNA/RNA, then a 260/280 nm absorbance measurement will be informative.
DEar Haitham Al-Madhagi
The simplest method is SDS-page and densitometric analisys of the band intensity.
At the following link you can find an example from my blog of densitometric analisys using the free imageJ software package
https://proteocool.blogspot.com/2021/09/tips-of-week-13-densitometric-protein.html
alternativelly you can use:
1) Capillary elettrophoresis, which require specific instumentation (
e.g;
https://www.perkinelmer.com/it/product/ht-labchip-gx-ii-touch-cls138160
https://sciex.com/products/capillary-electrophoresis/biophase-8800
2) Size exclusion chromatography as suggested from Sebastian Schmitt
(if you protein has single aggregation state). Since in SEC if your protein is a mix of monomer and dimer for example, you can obtain 2 peack from the same band observed in SDS-page, that is denaturing approach and dimer are broken.3) HPLC reverse phase chromatography (eg using a C18 coloumn)
good luck
Manuele
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