Antibody Labeling with Alexa Fluor 647

RN2N was covalently conjugated with Alexa Fluor 647 dye (Thermo Fisher Scientific) in PBS with 0.1 M sodium bicarbonate, as described previously^{14 (link)}. The labelled antibodies were then separated from free dye using a Superdex 200 10/300 column (GE Healthcare) equilibrated in 1 X PBS, pH 7.4, at 0.5 mL/min, and the degree of labeling was determined. Antibody-Alexa Fluor 647 conjugates were then concentrated to the desired injection volumes using an Amicon® Ultra filter (Millipore).

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Janowicz P.W., Leinenga G., Götz J, & Nisbet R.M. (2019). Ultrasound-mediated blood-brain barrier opening enhances delivery of therapeutically relevant formats of a tau-specific antibody. Scientific Reports, 9, 9255.

Publication 2019

Alexa Fluor 647 dye Superdex 200 10/300 column Amicon® Ultra filter

Alexa fluor 647 Antibody Sodium bicarbonate Ultra

Corresponding Organization :

Other organizations : Park Centre for Mental Health, University of Queensland

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Variable analysis

independent variables

Covalent conjugation of RN2N with Alexa Fluor 647 dye

dependent variables

Degree of labeling of antibody-Alexa Fluor 647 conjugates

control variables

Reaction carried out in PBS with 0.1 M sodium bicarbonate
Separation of labelled antibodies from free dye using Superdex 200 10/300 column equilibrated in 1 X PBS, pH 7.4, at 0.5 mL/min
Concentration of antibody-Alexa Fluor 647 conjugates to desired injection volumes using an Amicon® Ultra filter

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