PEROXIDASE-LABELED STICHOLYSINS METHOD FOR LOCALIZATION OF STICHOLYSINS IN THE SEA ANEMONE STICHODACTYLA HELIANTHUS.

Sticholysins I and II are the major cytolysins present in the venom of the sea anemone Stichodactyla helianthus. These polypeptides have been isolated, purified and characterized (Lanio et al. 2001), and their immunogenic and antigenic properties studied (Pico et al. 2000). Anemones were collected i...

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Publicado en:Revista VacciMonitor (Vacunología y Temas Afines) Vol. 11; no. 4; pp. 1 - 2
Autores principales: Pico, M. C., Noa, Y., Pérez, V., Izquierdo, M., Guerra, O., Infante, J. F., Otero, A.
Formato: Artículo
Publicado: Vacunas Finlay S.A. Oct2002
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Acceso en línea:Ver este registro en EBSCOhost
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        atl: PEROXIDASE-LABELED STICHOLYSINS METHOD FOR LOCALIZATION OF STICHOLYSINS IN THE SEA ANEMONE STICHODACTYLA HELIANTHUS.
      aug:
        au:
          Pico, M. C.
          Noa, Y.
          Pérez, V.
          Izquierdo, M.
          Guerra, O.
          Infante, J. F.
          Otero, A.
        affil:
          Centro de Estudio de Proteínas, Facultad de Biología, Universidad de La Habana. 25 No. 455 esq a J, Vedado, Ciudad Habana, Cuba
          Instituto Finlay. Ave 27 # 19807, Marianao, Ciudad Habana, Cuba
          Centro de Investigaciones de la Defensa Civil (CIDC). Carretera de Tapaste y 8 Vías, San José de las Lajas, Prov. Habana, Cuba
      su:
        Peroxidase
        Immunoenzyme technique
        Peptides
        Stichodactylidae
        Sea anemones
        Venom
      sug:
        subj:
          Peroxidase
          Immunoenzyme technique
          Peptides
          Stichodactylidae
          Sea anemones
          Venom
      ab: Sticholysins I and II are the major cytolysins present in the venom of the sea anemone Stichodactyla helianthus. These polypeptides have been isolated, purified and characterized (Lanio et al. 2001), and their immunogenic and antigenic properties studied (Pico et al. 2000). Anemones were collected in Miramar Coast, Ciudad Habana, fixed "in situ" in 4 % formaldehyde and 0.1 % glutaraldehyde, cut in small pieces and embedded in paraffin. Pieces were sliced to sections of 3 ?m, mounted on glass slides with polylysine, and then deparaffinized and rehydrated through xylene and a graded series of ethyl alcohol. To inhibit endogenous peroxidase sections were incubated for 10 min with 0.3 % hydrogen peroxide en methanol. Previously to immunohistochemistry stain samples were blocked 30 min with 1 % Triton X-100 in PBS and following incubated overnight with anti-sticholysins antibodies (0.25 ? g/mL) in PBS with 0.1 % Triton X-100 (PBS-TX). Afte r three 10 min rinses in PBS-TX the samples were incubated for 2 hours with peroxidase-labeled sticholysins diluted 1:500 in PBS-TX. Finally, the sections were washed 3 times and peroxidase activity was revealed by diaminobencidine. Control was performed replacing the anti-sticholysins by rabbit non-specific IgG. Using this method we can localize sticholysins in tentacles of anemone. The presence of the sticholysins in the tentacles suggests a role of these cytolysins in defense or prey capture processes of anemone. Lanio, M.E., Morera, V., Alvarez, C., Tejuca, M., Gómez, T., Pazos, F., Besada, V., Martínez, D., Huerta, V., Padrón, G. and Chávez, M.A. (2001) Purification and characterization of two hemolysins from Stichodactyla helianthus. Toxicon 39:187-194. Pico, M.C., del Monte, A., Calvo, L., Torres, Y., Felicó, E. and Otero, A.J. (2000) Inmunogenicidad y especificidad antigénica de las sticholisinas I y II de la anémona de mar Stichodactyla helianthus. Revista Biología 14(2):156-59.
      pubtype: Academic Journal
      doctype: Article
      src: R
    language: English
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