Optimize, Adapt and Overcome - 33

Optimize, Adapt, and Overcome: Tips from the Cell Line Development Front * Screening and Selection of GPCR-Expressing Cell Lines

Colonies picked by the ClonePix 2 System were
deposited into 96-well plates containing liquid
expansion media, cultured for one week, and
transferred to 384 well plates. Picked colonies
were functionally corroborated with the FLIPR
Tetra System using the FLIPR Calcium Assay 6 Kit.
The kit measures transient intracellular calcium
mobilization as a result of activation of the
M1 G-protein coupled IP3 sensitive pathway.
Carbachol was used as the muscarinic agonist.

A

C

B

Figure 2. (A) Selection of GPCR M1-expressing clones with both direct-labeled antibody
and (B) dual-labeled antibodies approaches shown (brightfield and fluorescence images).
The fluorescent intensity is proportional to M1 expression in the positive clones. (C) The
negative control, the CHO-K1 cell line, demonstrates no fluorescence in the PE channel.
33

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Optimize, Adapt and Overcome

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