Good afternoon,
I try to extract cell wall proteins from green macroalgae, I have tried different protocols and one of them included step of EGTA extraction, however this fraction is jelly like, so I expect polysaccharides from cell wall to be present. When loaded to SDS-PAGE, it was obvious that the gel pores were blocked and no bends were obtained, just light blue streak. According to Bradford, in this fraction I have quite a lot of proteins with which I would like to do Western Blot... I have already tried aceton precipitation, but still a lot of polysaccharides remained....So please do you have any recommendations how to get rid of these polysaccharides?
Thank you very much,
Tereza
Dear Tereza Přerovská
Pls. find the attached files
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4344596/
page 91 in: Andrew J. Hunt, James H. Clark. 2013. Element Recovery and Sustainability Royal Society of Chemistry,Pp.267. ISBN 1849736162, 9781849736169
http://www.nanodrop.com/Library/T111-Detection-and-Avoidance-of-Polysaccharides-in-Plant-Nucleic-Acid-Extractions.pdf
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-695X2013000400010
http://www.nature.com/pj/journal/v44/n8/full/pj2012105a.html
https://www.liquidwholefood.com/wp-content/uploads/2013/08/seaweedproteins.pdf
https://www.ncbi.nlm.nih.gov/pubmed/26108499
https://www.researchgate.net/post/What_is_the_best_way_to_precipitate_proteins_from_an_extract_which_also_has_polysaccharides
https://hal.archives-ouvertes.fr/hal-00107439/document
http://www.imedpub.com/articles/isolation-and-characterization-of-sulphated-polysaccharides-from-codiumtomentosum-j-stackhouse-1797-collected-from-southeast-coast.pdf
Hoping this will be helpful
Regards
Prof. Houda Kawas
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