Name: Arricchimento di proteine subcellulari in Leptospira utilizzando un approccio di frazionamento basato su Triton X-114 (Video) | JoVE Journal
Uploaded: 2025-08-08T12:30:00.000+00:00
Duration: 4 min 25 s
Description: Scopri l'arricchimento di proteine subcellulari in Leptospira mediante frazionamento con Triton X-114. Questo metodo consente di identificare fattori di virulenza nei batteri patogeni gram-negativi con elevata precisione.

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<li>Nogueira, S. V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Leptospira+interrogans+enolase+is+secreted+extracellularly+and+interacts+with+plasminogen%5BTitle%5D)+AND+%22PLoS+One%22%5BJournal%5D)">Leptospira interrogans enolase is secreted extracellularly and interacts with plasminogen.</a> PLoS One. 8 (10), e0078150(2013).</li>
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<li>Madathiparambil, M. G., Cattavarayane, S., Perumana, S. R., Manickam, G. D., Sehgal, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Presence+of+46+kDa+gelatinase+on+the+outer+membrane+of+Leptospira%5BTitle%5D)+AND+%22Curr+Microbiol%22%5BJournal%5D)">Presence of 46 kDa gelatinase on the outer membrane of Leptospira.</a> Curr Microbiol. 62 (5), 1478-1482 (2011).</li>
</ol>

<table><tbody><tr><td>Acido acetico</td><td>&nbsp; Sigma</td><td>&nbsp; 695092</td><td></td></tr><tr><td>Acetone</td><td>&nbsp; &nbsp; Sigma</td><td>&nbsp; 32201</td><td></td></tr><tr><td>Acrilamide</td><td>&nbsp; &nbsp; Sigma</td><td>&nbsp; A-3553</td><td></td></tr><tr><td>Persolfato di ammonio</td><td>&nbsp; &nbsp; Sigma</td><td>&nbsp; A-3678</td><td></td></tr><tr><td>Bilancia</td><td>&nbsp; SHIMADUZ</td><td>&nbsp; Z741079</td><td></td></tr><tr><td>Bis Acrilammide</td><td>&nbsp; &nbsp; Sigma</td><td>&nbsp; Codice: M7279</td><td></td></tr><tr><td>Frazione V dell'albumina sierica bovina (BSA)</td><td>&nbsp; Sigma</td><td>&nbsp; A-8022</td><td></td></tr><tr><td>Blu di bromofenolo</td><td>&nbsp; &nbsp; Sigma</td><td>&nbsp; B-8026</td><td></td></tr><tr><td>Provetta da centrifuga (50 mL)</td><td>Tarsons</td><td>500020</td><td></td></tr><tr><td>Pallone conico con tappo (150 mL)</td><td>Tarsons</td><td>431030</td><td></td></tr><tr><td>Centrifuga di raffreddamento</td><td>&nbsp; Eppendorf</td><td>&nbsp; 5355316200</td><td></td></tr><tr><td>Coomassie blu brillante</td><td>&nbsp; &nbsp; Sigma</td><td>1.12553</td><td></td></tr><tr><td>Microscopio a campo scuro</td><td>&nbsp; ZEISS</td><td>&nbsp; GmbH37081</td><td></td></tr><tr><td>Salviette delicate per attività</td><td>Kimtech Scienza</td><td>34155</td><td></td></tr><tr><td>Bagno secco</td><td>&nbsp; Eppendorf</td><td>&nbsp; 535531620</td><td></td></tr><tr><td>Ellinghausen&ndash; McCullough&ndash; Johnson&ndash; Harris (EMJH) medio, Difco</td><td>&nbsp; Difco</td><td>&nbsp; 279410</td><td></td></tr><tr><td>Vassoi per gel: i vassoi per gel devono essere di dimensioni larghe di almeno 2 cm e avere una superficie liscia e piatta con coperchio.</td><td>&nbsp; Hoefer</td><td>&nbsp; Non applicabile</td><td></td></tr><tr><td>Becco in vetro (150 mL)</td><td>Tarsons</td><td>421030</td><td></td></tr><tr><td>Glicina G8790</td><td>&nbsp; &nbsp; Sigma</td><td>&nbsp; G-8898</td><td></td></tr><tr><td>Carta da filtro assorbente Immobilon</td><td>Millipore</td><td>IBFP0813C</td><td></td></tr><tr><td>Impostazione semi-secca Immunoblot</td><td>&nbsp; ATTA</td><td>&nbsp; AE-6675</td><td></td></tr><tr><td>Incubatore - Per incubare i vassoi di gel per 18 ore a 37 °C.</td><td>&nbsp; EYELA</td><td>&nbsp; SLI-700</td><td></td></tr><tr><td>Flusso d'aria laminare</td><td>&nbsp; Non applicabile</td><td>&nbsp; Non applicabile</td><td></td></tr><tr><td>Luminol</td><td>&nbsp; Sigma</td><td>&nbsp; 09253</td><td></td></tr><tr><td>Cloruro di magnesio M8266</td><td>&nbsp; Sigma</td><td>&nbsp; Codice: M8266</td><td></td></tr><tr><td>Agitatore magnetico</td><td>&nbsp; TARSONI</td><td>&nbsp; 6040</td><td></td></tr><tr><td>Metanolo</td><td>&nbsp; &nbsp; Sigma</td><td>&nbsp; 17995-7</td><td></td></tr><tr><td>Puntali per micropipette (10 & micro; L&ndash; 10 ml)</td><td>Axygen</td><td>T-10, T-200-C, T-1000-C, T-10ML-C</td><td></td></tr><tr><td>Micropipette</td><td>&nbsp; Eppendorf</td><td>&nbsp; EP2231300010</td><td></td></tr><tr><td>Microprovette (1,5 mL Safe-Lock)</td><td>Eppendorf</td><td>EP022363212</td><td></td></tr><tr><td>Mini giro</td><td>&nbsp; Genaxy</td><td>&nbsp; GENERAZIONE MINI-6K</td><td></td></tr><tr><td>Lettore multimodale</td><td>BMG LABTECH</td><td>&nbsp; S/N 413-3877</td><td></td></tr><tr><td>Latte scremato in polvere, bovino</td><td>Sigma-Aldrich</td><td>Codice: M7409</td><td></td></tr><tr><td>Acido P-cumarico</td><td>&nbsp; &nbsp; Sigma</td><td>&nbsp; C9008</td><td></td></tr><tr><td>Soluzione salina tamponata con fosfato</td><td>&nbsp; &nbsp; Sigma</td><td>P4417&nbsp;</td><td></td></tr><tr><td>Frigorifero</td><td>&nbsp; LG</td><td>&nbsp; GC-B217BLJ2</td><td></td></tr><tr><td>Rocker - Agitatore oscillante o rotante a bassa velocità per colorare il gel.</td><td>&nbsp; TARSONI</td><td>&nbsp; 4080</td><td></td></tr><tr><td>SDS-PAGE SETUP - Apparecchio per elettroforesi</td><td>&nbsp; Hoefer, Inc.</td><td>&nbsp; SE300-10A-1.0</td><td>&nbsp; Modello Hoefer ' miniVE' che può colare gel di dimensioni 10 cm x 8 cm di spessore 1 mm.</td></tr><tr><td>Cloruro di sodio</td><td>&nbsp; &nbsp; &nbsp; Sigma</td><td>&nbsp; 5886</td><td></td></tr><tr><td>Dodecil solfato di sodio (SDS)</td><td>&nbsp; &nbsp; Sigma</td><td>&nbsp; L-4396</td><td></td></tr><tr><td>TEMED</td><td>&nbsp; &nbsp; Sigma</td><td>&nbsp; T-9281</td><td></td></tr><tr><td>Tritone X-114</td><td>&nbsp; &nbsp; Sigma</td><td>648468</td><td></td></tr><tr><td>Trizma Base (TRIS) T6066</td><td>&nbsp; &nbsp; Sigma</td><td>&nbsp; Codice: T6066</td><td></td></tr><tr><td>Vortice</td><td>&nbsp; TARSONI</td><td>&nbsp; 3020</td><td></td></tr></tbody></table>

arricchimento di proteine subcellulari in leptospira utilizzando un approccio di frazionamento basato su triton x-114

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Arricchimento di proteine subcellulari in Leptospira utilizzando un approccio di frazionamento basato su Triton X-114

Prokaryotes lack precise subcellular organelles but exhibit distinct regions such as cytoplasm, inner membrane, periplasm, and outer membrane, where most ...

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Arricchimento di proteine subcellulari in Leptospira utilizzando un approccio di frazionamento basato su Triton X-114

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