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Methods Mol Biol ; 2180: 271-283, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-32797415

RESUMO

Ice recrystallization inhibition assays are used to screen for compounds that possess the ability to inhibit ice recrystallization. The most common of these assays are the splat cooling assay (SCA) and sucrose sandwich assay (SSA). These two assays possess similarities; however, they vary in their sample size, cooling rate, and the solution used to dissolve the analyte. In this chapter, both assay methods are described in detail, and we perform a direct comparison of the assays by evaluating the IRI activity of an antifreeze protein (AFP I). IRI activity is quantified by using ImageJ software to analyze ice crystals, and a quantitative value describing the efficiency of the inhibitor is generated. This analysis emphasizes the importance of choosing the right assay to measure IRI activity.


Assuntos
Proteínas Anticongelantes/metabolismo , Criopreservação/métodos , Crioprotetores/farmacologia , Congelamento , Ensaios de Triagem em Larga Escala/métodos , Gelo/análise , Animais , Bioensaio , Cristalização , Humanos , Transição de Fase
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