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en:aree:epr:altre_risorse:biacore:metodica [2024/04/18 20:24] Gianluca Frustagli Changes due to move from Proteomics Area to EPR Area. |
en:aree:epr:altre_risorse:biacore:metodica [2024/04/19 19:52] Gianluca Frustagli [The Sensogram] Fixed link. |
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| {{ :aree:epr:altre_risorse:biacore:absolute_response.png |}} | {{ :aree:epr:altre_risorse:biacore:absolute_response.png |}} | ||
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| - | Dopo un certo tempo di perfusione del campione, il sistema effettua un lavaggio con il //running buffer// durante il quale si ha il distacco delle molecole di analita che hanno interagito con il ligando. Infine, viene flussato un //tampone rigenerante// tipicamente molto salino e basico che consente di staccare tutte le molecole di analita dal ligando, rendendo disponibile il chip ad un nuovo ciclo di legame-distacco. | ||
| After sample injections, molecules of the sample, buffer ions and impurities start interacting with the sensor surface until there is full saturation of the sites, parts of these interactions are not specific, thus just after there is washing out of the fluidic system by running buffer, the detachment of unspecific interactions occurs leaving the real interaction against the baseline. Moreover, according to the experiment, it is also possible to choose two alternative modalities: consecutive injections vs repetitive | After sample injections, molecules of the sample, buffer ions and impurities start interacting with the sensor surface until there is full saturation of the sites, parts of these interactions are not specific, thus just after there is washing out of the fluidic system by running buffer, the detachment of unspecific interactions occurs leaving the real interaction against the baseline. Moreover, according to the experiment, it is also possible to choose two alternative modalities: consecutive injections vs repetitive | ||
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| From the Sensogram it is possible to obtain kinetics information such as association and dissociation by fitting experimental data with specific models as well as thermodynamic information (enthalpy, entropy and Free Gibbs energy) since the device can control the temperature from 4 to 45 °C. \\ | From the Sensogram it is possible to obtain kinetics information such as association and dissociation by fitting experimental data with specific models as well as thermodynamic information (enthalpy, entropy and Free Gibbs energy) since the device can control the temperature from 4 to 45 °C. \\ | ||
| - | Amongst the //[[en:aree:proteomica:biacore:applicazioni|applications]]// are presented some explanatory approaches to catch these parameters. | + | Amongst the //[[en:aree:epr:altre_risorse:biacore:applicazioni|applications]]// are presented some explanatory approaches to catch these parameters. |
