Channels

 

Special Offers & Promotions

 

 

Latest News

 

 

View Channel

New Products

 

 

View Channel

Video Presentations

 

 

View Channel

Separation Science

 

 

View Channel

Microscopy & Image Analysis

 

 

View Channel

Laboratory Automation & IT Solutions

 

 

View Channel

 

Quantifying Receptor-Ligand Binding to GPCRs in Live Cells - A new HTS format Replaces Radioactive Ligand Studies

PHERAstar FS microplate reader from BMG LABTECH for HTS and Core Lab facilities

Ligand binding affinities at G-protein coupled receptors (GPCRs) have historically been determined using a radioligand that competes for receptor binding sites against an unlabeled drug-like compound. But the potential hazards of open-source radioisotope handling, and the environmental impact of radioisotope disposal, make this a less desirable, costly technology


Fluorescent Ligand Binding Assays in Live Cells Use High Content Analysis (HCA)
New methods, such as the development of fluorescent ligands for GPCRs (CellAura Tehnologies, Ltd.) provide a safer method for determining ligand binding affinities. However, measurement of fluorescent ligands binding to cells relies on capturing and processing complex high resolution images. This approach can be lengthy and can result in 96-well plate read times of approximately 30 minutes or more. Cell-based assays that take too long can undergo changes that could affect the assay results. Therefore, these High Content Analysis (HCA) assays are too slow to be useful for High Throughput Screening (HTS).

HTS Microplate Reader is Sensitive Enough for Live Cell Assays
Here enters the PHERAstar FS microplate reader from BMG LABTECH for HTS and Core Lab facilities. Using the adjustable focal height and advanced bottom reading features on the PHERAstar FS, live cells are quickly and easily quantified for fluorescent ligand binding to the adherent cell layer in a 96-well plate format. Depending on several variables, a 'read to IC50 pKi curve plot' times were between 2.5 and 10 minutes for a 96-well plate. This is in stark contrast to a similar HCA assay, which can take between 30-60 minutes depending on the measurement variables.

Here is example data outlined in the application note Quantifying Fluorescent Ligand Binding to GPCRs in Live Cells using the PHERAstar FS - a new format for HTS. The PHERAstar FS performs GPCR ligand binding assays using two fluorescent ligands from CellAura for adenosine (A1 and A3) and dopamine (D1) receptors. Derived pKi curve plots were comparable to previous radioligand binding assays and good Z’ values show that these fluorescent binding assays with this microplate reader are amendable for high throughput screening.

Safer, Faster Receptor-Ligand Binding Assays
The PHERAstar FS and CellAura fluorescent ligands provide a rapid and robust way to quantify receptor-ligand binding in live cells. Using a 96-well format and a simple 'add > mix > wash > read' assay protocol, whole-cell radioligand binding assays are replaced with this safer and more cost effective alternative. In addition, the time saving advantages over HCA assays make this format suitable for high throughput screening (HTS). In addition, multiplexing with other fluorophores can be used to normalize for cell number such (i.e. Hoechst or DAPI) or to measure downstream second messenger effects of like calcium flux or cAMP production.

Find out more about fluorescent ligands at CellAura Technologies Ltd.

To find out more about the PHERAstar FS HTS microplate reader please visit www.bmglabtech.com.


If you have not logged into the website then please enter your details below.



 

News Channels

 

 

Subscribe to any of our newsletters for the latest on new laboratory products, industry news, case studies and much more!

Newsletters from Lab Bulletin

 

Request your free copies HERE

 

 

 

Popular this Month

Top 10 most popular articles this month

 

 

Today's Picks

 

 

 

 

Looking for a Supplier?

Search by company or by product

 


Company Name:

Product:


 

 

 

 

Please note Lab Bulletin does not sell, supply any of the products featured on this website. If you have an enquiry, please use the contact form below the article or company profile and we will send your request to the supplier so that they can contact you directly.

Lab Bulletin is published by newleaf marketing communications ltd.


 

Media Partners

 

Exhibitions & Events