I am doing the estimation of protein of marine microalgae but unable to get proper reading for the standard curve using BSA as well as sample. I just want to know about the extraction procedure for protein.
I do not have "a protocol" (sorry) but it is a well known problem...
Give this link a try
http://www.ncbi.nlm.nih.gov/pubmed/22138185
Good luck.
Yoram
Collect an ample amount of micoalgal cell by centrifugation.
Homogenized in a protein extraction buffer
Precipitate the protein
Collect the protein via centrifugation, wash with ethanol in tris buffer
Determine protein by Bradford method
A cause of discrepancies can be the choice of empirical formula for microalgal protein. The following paper may be of help:
Heaven S, Milledge J, and Zhang Y (2011) Comments on 'Anaerobic digestion of microalgae as a necessary step to make microalgal biodiesel sustainable'. Biotechnology Advances 29, 164-7.
The following reference may be of help:
Laurens LML, Dempster TA, Jones HDT, Wolfrum EJ, Van Wychen S, McAllister JSP et al. (2012) Algal Biomass Constituent Analysis: Method Uncertainties and Investigation of the Underlying Measuring Chemistries. Analytical Chemistry 84, 1879-87.
@priti; if you are not getting proper standard curve then its a big fault you are doing during procedure because no extraction is needed for standard curve. I think your chemicals are contaminated. change them. or may be you are not preparing the buffer well. pH is the main factor during buffer preparation.
For extraction of sample you can use many ways.e.g. hot extraction using NaCl, with pistle and mortar using buffer, and even you can sonicate your material to break the algal cell.
Tell what protein estimation method you are using? Lowry or Bradford?
- Badges
- Science topic
- Similar topics
- Mathematics