Solutions of DesAb-O (4 and 2 μM) were spotted (3.5 μL) on a 0.2-μm-pore-size nitrocellulose membrane (GE Healthcare). As negative control, samples with same amounts of DesAb-PYGSMYVHS, which showed the weakest binding to Aβ40 oligomers in in the ANS binding measurements, were used. The blots were blocked in PBS and 5% BSA overnight at 4 °C. Then, they were incubated in solutions containing 5 μM Aβ40 oligomers in PBS overnight at 4 °C. Blots with bound oligomers were then probed with the mouse monoclonal anti-Aβ antibody [6E10] (1:2,000 dilution; Absolute Antibody Ltd) and with goat anti-mouse IgG (H+L) secondary antibody and AF488 conjugate (1:5,000 dilution; Life Technologies).
Goat anti mouse igg h l secondary antibody and af488 conjugate
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
Goat anti-mouse IgG (H+L) secondary antibody and AF488 conjugate is a laboratory reagent used for the detection and quantification of mouse immunoglobulin (IgG) in various immunoassays and immunohistochemical applications. It is a polyclonal antibody raised in goats against the heavy and light chains of mouse IgG, and is conjugated to the Alexa Fluor 488 fluorescent dye.
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Lab products found in correlation
2 protocols using goat anti mouse igg h l secondary antibody and af488 conjugate
1
Binding of Aβ40 Oligomers to DesAb-O
2
Dot Blot Analysis of DesAb Binding
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In order to assess the binding of the DesAbs, dot blots were prepared as follows: 4 μM solutions of DesAb were spotted (3.5 μL) onto a 0.2 μm pore size nitrocellulose membrane corresponding to total deposited antibody of 50, 25, 12.5 and 6.3 µg DesAb. The blots were blocked in PBS, 5% BSA overnight at 4 °C. Subsequently, they were incubated in solutions containing 5 μM Aβ40 oligomers in PBS overnight at 4 °C. Blots were then probed with the mouse monoclonal anti-amyloid β antibody (clone 6E10) (1:2000 dilution; Absolute Antibody Ltd., Oxford, UK) overnight at 4 °C, and then with goat anti-mouse IgG (H+L) secondary antibody and AF488 conjugate (1:5000 dilution; Life Technologies) for 2 h at 25 °C.
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