A fast centrifuge method for nematocyst isolation from Pelagia noctiluca Forskal (Cnidaria: Scyphozoa).
Nematocyst isolation from surrounding tissue is an important step to characterize Cnidarian venom. Although several protocols have been used to extract venoms from cnidarian tissues, the complete isolation of nematocysts from tissue is still difficult. The goal of the present work was to evaluate the effectiveness of three different media, Percoll, Ficoll and Methylcellulose in isolating nematocysts from Pelagia noctiluca tentacles by centrifugation. The complete sedimentation of nematocysts and tissue fragments to the bottom of the test tubes was observed in Ficoll and Methylcellulose suspensions. The best result was obtained using a discontinuous density gradient of Percoll: three types of nematocysts were concentrated in three different fractions along the density gradient. Protein assay and preliminary chromatographic analyses confirmed these results.
Marchini
Barbara
B
Dipartimento di Medicina Interna, Università di Pisa, Italy.
De Nuccio
Luca
L
Mazzei
Mauro
M
Mariottini
Gian Luigi
GL
D000825:Animal Structures / Q000737:chemistry; D000818:Animals; D002458:Cell Fractionation / Q000379:methods; D002469:Cell Separation / Q000379:methods; D002499:Centrifugation, Density Gradient / Q000379:methods*; D002853:Chromatography, Liquid; D003064:Cnidarian Venoms / Q000302:isolation & purification*; D005362:Ficoll; D008747:Methylcellulose; D011205:Povidone; D037401:Scyphozoa / Q000737:chemistry / Q000648:ultrastructure*; D012822:Silicon Dioxide; D013056:Spectrophotometry, Ultraviolet
D003064:Cnidarian Venoms; D005362:Ficoll; C016039:Percoll; D012822:Silicon Dioxide; D008747:Methylcellulose; D011205:Povidone