I am new to protein chemistry,
Can I know how do I check oligomers / dimers of the protein using Native Gel ? Thanks in advance.
Hi, I agree with Dylan. Size exclusion chromatography only separates proteins based on size (if your protein is globular). Native PAGE separates proteins based on size and charge.
Run ur sample in non reducing condition on Sds gel with the known marker , in case of oligomer or dimer protein band will run less than monomer unit, after this u can run the protein in reducing condition if u see 2 bands than it will be oligomer and if u see a single band than it must be the dimer
Hi
thanks for suggestions.
Can I undersand whether you mean run SDS gel with DTT and without DTT ? I did this already. But I couldnt find any differnce in the gel band.
Is there any other way apart from Gel filtration / MALS to determine dimerisation of the protein ? Thanks in advance
Yes, i do mean the same... in case you are having a pure sample you can digest the protein and look into the MS/MS profile. If you see peptides of the same proteins you can confirm the purity of the compound, secondly i was curious what made you put an assumption of your protein to be oligomer/dimer... is there any disulfide bonds in the protien
The best way to check for this kind of protein configuration is using a calibrated size exclusion column. You'll be able to determine the number of subunits in your protein, assuming the column resolution is calibrated in the region where your protein elutes.
Hi, I agree with Dylan. Size exclusion chromatography only separates proteins based on size (if your protein is globular). Native PAGE separates proteins based on size and charge.
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