GABAB (B1a/B2) Human GABAB GPCR Binding Antagonist Radioligand LeadHunter Assay - TW

Procedure Summary:

Human recombinant GABAB(1a) receptors expressed in CHO-K1 cells are used in modified Tris-HCl buffer pH 7.4. A 1 μg‡ aliquot is incubated with 4 nM [³H]CGP-54626 for 180 minutes at 25℃. Non-specific binding is estimated in the presence of 3000 μM GABA. Receptors are filtered and washed, the filters are then counted to determine [³H]CGP-54626 specifically bound. Compounds are screened at 10 μM.
Note: ‡Membrane protein may change from lot to lot, the concentration used will be adjusted if necessary.

Ligand:

[³H] CGP-54626

Ligand Kd (nM):

3.3

Ligand Concentration:

4 nM

Non Specific:

3000 µM GABA

Incubation:

180 min at 25°C

Control Inhibitor:

Cgp-54626

Criteria For Significance:

≥ 50% of max stimulation or inhibition

Test Concentration / Dose:

Suggested concentration for initial testing is 10 uM

Test Sample Requirements:

Minimum for 1) Screen: 25 μl of 10 mM stock -OR- 1 mg (pre-weighed). 2) Dose Response: 60 μl of 10 mM stock -OR- 1 mg (pre-weighed).

Minimum Order Quantity:

2