Dear Friends,
I am naive in fielf of Proteomics, I have been searching a protocol for isolation of protein from plant root and shoots. I want to do Protein estimation from isolated proteins.
If anyone out there has gone through this process or tried to, your advice would be greatly appreciated.
There are many extraction protocols for plant roots and shoots, including TCA/acetone precipitation, phenol method, urea lysis buffer, and combination of TCA/acetone and phenol method. Unfortunately there is no universal extraction method for plant proteins. Each plant may contains different level of interfering contaminants. Therefore, that is good to optimize which protocol works the best for your plant sample. TCA/acetone precipitation is the best start as it just involves a few consumables and easy to perform. However, the too dried pellets might be difficult to solubilize in your buffer. You need to be more careful on this step. Phenol method works well for many plant species but phenol is toxic. Shoots may contain more interfering compounds compared to roots.
For measuring the protein content, you may use commercially available kits or conventional methods such as Bradford, Lowry, etc.
As stated above there are many ways to extract proteins, if your main goal is just to estimate protein concentration then any protocol would do (through ionic or organic or isoelectric point) and and estimation assays will work.. but if you want your protein in native state an be viable for any further tests you will need to do the extraction in an ionic solution or even in a water... (usually maximum O.D. readings are from water extract).
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