receptor cloning - screening clones for expression of receptors (Jul/08/2005 )
I am interested in stable transfection of human receptor gene in mammalian cell line. I have been looking at literature for the same.
Our lab do not have a prior expereince on these type of studies. In the literature i have come across screening methods like ELISA or western blot.
What are the commonly used techniques for screening clones for receptor expression?
I am not very sure of how to manage hundreds of clones in 96 well plate, how to characterise them, or screen for best possible expressor clone.
Out of ELISA and western blot, which one will be an easier technique for screening?
can any one please guide me and provide suggestions?
thanks a lot.
Elisa is the easier way to screen lots of clones if you have an elisa up and running and is reliable.
If not, well, I hope you have a good antibody to screen by western blot...
Is there anyway you can screen by activity assay?
Thanks for ur guidance.
Western blot control (receptor transfected cell postive control lysate) and receptor antibody for western blot application are commercially available.
Receptor antibody for ELISA is also available, but receptor peptide (blocking peptide) to be used for ELISA standard is not available.
In this scenario, is there any possibility to develop ELISA for screening?
Eventually 1 or 2 lead clones will be used for development of receptor radioligand binding assay.