Hello Every one
My purified (single peak in HPLC) sample is glycoprotein (3.5 microgram/100 microgram ) with molecular weight range between 60-80 kDa using SDS PAGE. I had given my sample two times in separate places for MALDI-TOF using sinnapinic acid as matrix but they told it is not ionizing.Please suggest me how to know Molecular weight of this glycoprotein.
If the glycosylation is N-liked, you can remove it with the enzyme PNGase F. After that treatment, when you run it on SDS-PAGE you will see a narrower band with the correct molecular weight of the polypeptide chain.
Thanks a lot for your suggestion. But we cant detect using MALDI-TOF.
Is sinapinic acid the only matrix that was suggested? There may be some other matrix that works better in MALDI-ToF.
To what precision do you need the answer?
If a good estimate of solution Mwt is needed, suggest light scattering or analytical ultracentrifugation (will give a answer to within 5%). For more accuracy, you may be able to get it by nanospray Mass spec as well.
@Zohara Nafeesa Did you quantify the glycoprotein?? if yes which method you follows.
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