I am dealing with a lectin which is capable of agglutinating only neuraminidase treated RBCs and not trypsinized RBCs. Can I get a working protocol for removal of sialic acid residues from RBC surface proteins?
Neuraminidase (25 mU/mL, from Arthrobacer ureafaciens) was added to washed RBC (5 × 107 cells/mL) and incubated for 30 min at 37 °C to remove the sialic acid, reaction was stopped by three washing with ice cold Hepes buffered saline (HBS), removing and adding equal volumes of HBS.
Thank you Gregory, for the suggestion. However, the neuraminidase we have in our lab is from Clostridium perfringens. Will the same conc. of my neuraminidase be effective? Please advise.
Dear Suhas,. I have no experience with Clostridium. I can only advise you to check relevant articles on the Internet and Pubmed. For example: http://vir.sgmjournals.org/content/70/12/3347.full.pdf
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