Antibody binding was determined using a FortéBio Bio-Layer Interferometry instrument (Sartorius Octet Red96e) at 25°C with a shake speed of 1000 rpm. Antibodies were diluted to 20 μg/mL in a flat bottom 96-well plate (Greiner) with .22μm filtered phosphate buffered saline pH 7.4 and 0.05% Tween 20 (PBS-T). The antigens were diluted to a concentration of 50μg/mL using PBS-T. Hydrated Anti-hIgG Fc Capture (AHC) biosensors (Sartorius #18–5060) were equilibrated for 60 second and then antibodies were loaded to biosensors for 300 seconds. After a 60-second wash and a 180-second baseline step, biosensors were then dipped into the diluted antigens for a 200-second association. Next, antibody and antigens allowed to dissociate for 300 seconds. Data was analyzed using Data Analysis HT 12.0 software. The negative control antibody, CH65, was indicated as a reference sensor and subtracted from the remaining ligand sensor measurements. Data was then aligned to the average of the baseline step and plotted using GraphPad Prism 9 software.
Bio layer interferometry instrument
Manufactured by Sartorius
The Bio-Layer Interferometry instrument is a label-free, real-time detection technology that measures binding interactions between molecules. It uses optical biosensing to monitor the interference pattern of white light reflected from a sensor surface to detect and quantify biomolecular interactions.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using bio layer interferometry instrument
1
Antibody Binding Kinetics Analysis
2
Antibody-Antigen Binding Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Antibody binding was determined using a FortéBio Bio-Layer Interferometry instrument (Sartorius Octet Red96e) at 25°C with a shake speed of 1000 rpm. Antibodies were diluted to 20 μg/mL in a flat bottom 96-well plate (Greiner) with 0.22 μm filtered phosphate buffered saline pH 7.4 and 0.05% Tween 20 (PBS-T). The antigens were diluted to a concentration of 50 mg/mL using PBS-T. Hydrated Anti-hIgG Fc Capture (AHC) biosensors (Sartorius #18–5060) were equilibrated for 60 s and then antibodies were loaded to biosensors for 300 s. After a 60-s wash and a 180-s baseline step, biosensors were then dipped into the diluted antigens for a 200-s association. Next, antibody and antigens allowed to dissociate for 300 s. Data was analyzed using Data Analysis HT 12.0 software. The negative control antibody, CH65, was indicated as a reference sensor and subtracted from the remaining ligand sensor measurements. Data was then aligned to the average of the baseline step and plotted using GraphPad Prism 9 software.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!