RV: Lectin agglutination of promastigotes

[Javier Moreno]

We are trying to separate procyclic from metacyclic promastigotes of L. infantum by agglutination of the procyclics with PNA (SIGMA). The promastigotes are from a young culture (3rd subculture) growing in RPMI. Till now lectin concentrations of up to 500 microgr./ml (in PBS) have had no visible effect (the promastigotes (10 millions/ml) were previously washed 4 x in PBS and the incubation lasted 1h at 23ºC.) What's wrong?
If anybody has experience in handling lectins and would be so nice to help me I would be VERY grateful. If somebody has a better idea how to separate the pro- from the meta-cyclic promastigotes then don't hesitate to send me a message. Thanks a lot,
					Jean Marc Lohse
					Dept. Parasitology
					Centro Nacional de Microbiología
					Madrid