Differential analysis of cell surface proteins on closed...

G - Physics – 01 – N

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G01N 33/50 (2006.01) C07K 1/13 (2006.01) G01N 33/58 (2006.01)

Patent

CA 2525685

Disclosed are matched fluorescent reagents and a method for reproducibly labelling membrane components, such as those expressed on the cell surface, and subsequent differential analysis of the labelled components to detect differences between cell types and states. Furthermore, the present method utilises an internal standard in order to match protein patterns across gels thereby avoiding gel-to-gel variation. The method according to the invention is particularly useful, for example, for detecting low abundance membrane proteins, for detecting changes in receptors expressed in the cell membrane, for example on ligand binding, or in response to stimuli.

L'invention concerne des réactifs fluorescents appariés ainsi qu'un procédé de marquage reproductible de composants membranaires, tels que ceux exprimés à la surface cellulaire, et d'analyse différentielle ultérieure des composants marqués pour détecter les différences entre les états et les types de cellules. Par ailleurs, ce procédé fait intervenir un étalon interne pour apparier les motifs protéiques à travers les gels et éviter la variation gel-gel. Selon l'invention, ce procédé s'utilise particulièrement, par exemple, pour détecter des protéines membranaires de faible abondance, pour détecter les modifications des récepteurs exprimés sur la membrane cellulaire, par exemple sur une liaison aux ligands, ou en réponse à un stimuli.

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