p51_FLAG was diluted to 8 μM before preparing a 2-fold dilution series down to 0.16 μM. A3G was purified as below, dialyzed against PBS + 0.01% IGEPAL CA 630 and used at a final concentration of 10 nM. Interaction analyses were performed on a Biacore T200 instrument (GE Healthcare). Binding surfaces for 6xHis tagged A3G were created by immobilizing an anti-His antibody (Biacore/GE Healthcare) onto a Series S CM5 sensor chip surface (Biacore/GE Healthcare) by amine coupling. The capture of the A3G ligand was carried out at low surface density (~200 RU) to minimize potential A3G-A3G interactions and ensure monomeric interactions with p51. The p51 analyte was injected at different concentrations, each in duplicate, in running buffer (PBS + 0.01% IGEPAL CA 630) at 15 μl/min for 4 min followed by a 15 min dissociation time. Standard double referencing data subtraction methods were applied9 (link).
Series s cm5 sensor chip surface
Manufactured by Cytiva
The Series S CM5 sensor chip surface is a laboratory equipment product designed for use in surface plasmon resonance (SPR) analysis. It serves as a solid support for the immobilization of various biomolecules, enabling the study of molecular interactions in real-time.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using series s cm5 sensor chip surface
1
Quantifying p51 Binding to A3G Enzyme
2
Quantifying p51 Binding to A3G Enzyme
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p51_FLAG was diluted to 8 μM before preparing a 2-fold dilution series down to 0.16 μM. A3G was purified as below, dialyzed against PBS + 0.01% IGEPAL CA 630 and used at a final concentration of 10 nM. Interaction analyses were performed on a Biacore T200 instrument (GE Healthcare). Binding surfaces for 6xHis tagged A3G were created by immobilizing an anti-His antibody (Biacore/GE Healthcare) onto a Series S CM5 sensor chip surface (Biacore/GE Healthcare) by amine coupling. The capture of the A3G ligand was carried out at low surface density (~200 RU) to minimize potential A3G-A3G interactions and ensure monomeric interactions with p51. The p51 analyte was injected at different concentrations, each in duplicate, in running buffer (PBS + 0.01% IGEPAL CA 630) at 15 μl/min for 4 min followed by a 15 min dissociation time. Standard double referencing data subtraction methods were applied9 (link).
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