I am working on seed storage proteins, where I am extracting whole protein using Urea buffer. please suggest me best quantification method where I want to run the sample in 2D.
What you choose will depend on how much sample you have and what is available to you. The best method is amino acid analysis. More likely to be available is simple A280, ideally you'll have access to a Nanodrop, or something similar, as this allows for low volumes. Just make sure you choose an appropriate blank.
Hi there,
To go further in Peter's way: the appropriate blank being the buffer alone (7M Urea, 2M Thiourea, 30mM Tris). What I always do is to set the blank and immediately to run it as a sample: if the blank is OK then the spectrum is totally flat and null. If not then redo the blank until it's flat.
If you have enough sample to use a small portion of it for a protein assay, then you can use the Pierce™ 660nm Protein Assay Reagent from Thermo-Fisher. The sample will have to be diluted a few-fold to get the thiourea and urea concentrations into an acceptable range. Here is the web page for the product:
https://www.thermofisher.com/order/catalog/product/22660
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