Biotinylated-BSA (1 μg/μl, Thermo) is flowed for 25 min through the device, allowing its binding to the epoxy surface. On top of the biotinylated-BSA, 0.5 μg/μl of Neutravidin (Pierce, Rockford, IL) is added (flow for 20 min). The 'button' valve is then closed, and biotinylated-PEG (1 μg/μl, (PG2-AMBN-5k, Nanocs Inc.) is flowed over for 20 min, passivating the flow layer, except for the buttons area. Following passivation, the 'button' valve is released and a flow of 0.2 μg/μl biotinylated anti-GFP antibodies (Abcam; #ab6658, Cambridge, United Kingdom) or 0.01 µg/µl biotinylated anti-Flag antibodies (Cell Signaling; #2908S Danvers, MA, USA) were applied. The antibodies bound to the exposed Neutravidin, specifically to the area under the 'button', creating an array of anti-GFP -or anti-Flag tag. PBS buffer was used for washing in between steps. In the case of p27 immobilization, surface chemistry was performed with 0.2 μg/ml donkey anti-mouse whole IgG antibodies (#715-065-150, Jackson Immuno research laboratories, Maryland, USA) followed by 20 min flow of 6.5 μg/ml anti p27 antibodies (Santa Cruz biotechnology, Heidelberg Germany; #1641 mouse).
Pg2 ambn 5k
Manufactured by Nanocs
The PG2-AMBN-5k is a laboratory equipment product. It is a specialized device designed for specific laboratory applications. Without further details about the intended use or technical specifications, a concise and unbiased description cannot be provided while maintaining objectivity. More information would be required to create a detailed and accurate description of the product's core function.
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2 protocols using pg2 ambn 5k
1
Microfluidic Antibody Immobilization
2
Surface Bioconjugation Biomolecule Immobilization
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Biotinylated-BSA (1 μg/μl, Thermo) is flowed for 25 min through the device, allowing its binding to the epoxy surface. On top of the biotinylated-BSA, 0.5 μg/μl of NutraAvidin (Pierce, Rockford, IL) is added (flow for 20 min). The ‘button’ valve is then closed, and biotinylated-PEG (1 μg/μl, (PG2-AMBN-5k, Nanocs Inc.) is flowed over for 20 min, passivating the flow layer, except for the buttons area. Following passivation, the ‘button’ valve is released and a flow of 0.2 μg/μl biotinylated anti-GFP antibodies (Abcam; #ab6658, Cambridge, United Kingdom) or 0.01 µg/µl biotinylated anti-Flag antibodies (Cell Signaling; #2908 S Danvers, MA, USA) were applied. The antibodies bound to the exposed NutraAvidin, specifically to the area under the ‘button’, creating an array of anti-GFP - or anti-Flag tag. PBS buffer was used for washing in between steps. In the case of p27 immobilization, surface chemistry was performed with 0.2 μg/ml donkey anti-mouse whole biotinylated anti-IgG antibodies (#715-065-150, Jackson Immuno research laboratories, Maryland, USA) followed by 20 min flow of 6.5 μg/ml anti p27 antibodies (Santa Cruz biotechnology, Heidelberg Germany; #1641 mouse).
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