MCH1 Human Melanin-Conc. Hormone GPCR Binding Agonist Radioligand LeadHunter Assay - TW

Procedure Summary:

Human recombinant recombinant melanin-concentrating hormone MCH1 (SLC1) receptors expressed in CHO-K1 cells are used in modified HEPES buffer pH 7.4. A 0.08 μg‡ aliquot is incubated with 0.05 nM [¹²⁵I]-Tyr-S36057 for 120 minutes at 25℃. Non-specific binding is estimated in the presence of 1 μM MCH (human, mouse, rat). Membranes are filtered and washed, the filters are then counted to determine [¹²⁵I]-Tyr-S36057 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:

[¹²⁵I] Tyr-S36057

Ligand Kd (nM):

0.1

Ligand Concentration:

0.05 nM

Non Specific:

1 µM MCH (human, mouse, rat)

Incubation:

120 min at 25°C

Control Inhibitor:

MCH (human, mouse, rat)

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