I study the lectin activity of purified lectins from Linum anthers and pistils. The extractant composition includes a detergent Triton X-100 and the lectin activity is determined by hemagglutination assay. In the presence of detergent occurs the hemolysis of erytrocytes. How I can remove this detergent from solution?
Hi, you can freeze-dry an use crystals of lectins. In next purification, do not use triton more than 0.02 percent.
Dear Arash,
Thank you! In future I will use 0,02 % of this detergent.
Best regards,
Hanna
Firstly, you do not any detergent for extraction but if you already has it in your extract, try dialysis against saline solution (0.9% NaCl) or ultradiafiltration. Currently plant lectins have molecular weights above 20-30 kDa it is easy to find the right cut-off membranes. Look for papers 1980-1990 for plant lectins extraction methods, several researchers were working with plant lectins this period. Personally, I have done good extraction with just 0.9% NaCl.
Dear Ana Maria,
Thank you!
I used detergent because I need extracted not only soluble lectins. I need extracted membrane and cell wall lectins...
Best regards,
Hanna
Possibly there is the same lectin into de different tissues of the plant. Try firstly with 0.9% NaCl and once the lectin be extracted, assayed and identified or characterize, try other variants. Anyway, detergents are removed by dialysis or ultradiafiltration with a lot of wash changes. Alternatively, use sonication for cell wall proteins extraction.
I see you have already extracted and identified linum lectin ¿or lectins?. Therefore, I think you are doing the re-extraction of the residue after saline extraction with detergent, isn't it?. Try dialysis for removing detergent. Also, if you try your erythrocytes with glutaraldehyde or glycerol (I do not remember well) they resist detergent.
Yes, I am doing the re-extraction of the residue after saline extraction with detergent. Thank you - I will try used erythrocytes with glycerol (I use rabbit erythrocytes and the glutaraldehyde is destroys this red blood cells).
To remove detergent put the extract into a high salt (1 M or greater) solution and dialyze exhaustively. It could be repeated, but probably does not need the repeat.
- Badges
- Science topic
- Similar topics
- Analytical Chemistry
- Extraction