NAN-190 Fluorescent ligand (Red) (ab118177)
- Product nameNAN-190 Fluorescent ligand (Red)
Fluorescent 5-HT1A antagonist
- Biological descriptionFluorescent 5-HT1A antagonist (pKD values are 8.75 and 6.34 at 5-HT1A and 5-HT2A respectively). Wide range of applications which include localizing receptor distribution in tissues and cells, and live-cell imaging of receptor kinetics.
Excitation wavelength: 633 nm
Emission wavelength: 650 nm
- Purity> 97%
- Selectivity5-HT1A: 8.75 | 5-HT1B: 5.57 | 5-HT2A: 6.34
- Molecular Weight1025.00
- Molecular formulaC55H59N8O7F2BS
- Validation notesCells were plated in clear bottom black 96-well tissue culture plates and grown to confluence for 24 hours. Medium was then removed and cells incubated in loading buffer comprising HEPES buffered saline with 0.5 mM Brilliant Black BN, 2.5mM probenecid, 0.023% pluronic acid F-127 and 2.5ug/ml Fura-2 AM at 37ºC for 30 – 45 mins, either in the presence of ab118177 or a DMSO vehicle control. During this time serial dilutions of the 5-HT1A agonist, 5-HT, were prepared in HEPES buffered saline containing 2.5 mM probenecid and 0.5 mM Brilliant Black BN.
To determine the antagonist activity of ab118177, serial dilutions of the agonist was added to wells with or without ab118177. The agonist Ca++ response in the absence or presence of ab118177 was determined by exciting at 340 nm and 380 nm and ratioing the fluorescence intensity of Fura-2 signal collected at 320 nm.
The KD was calculated from the shift of the agonist response curve.
- Storage instructionsStore at -20°C. Avoid exposure to light.
- HandlingFor imaging at the 5-HT1A receptor use solutions up to 200 nM in DMSO.
Our Abpromise guarantee covers the use of ab118177 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Fluorescent Cell Imaging||Fluorescent Cell Imaging: Use at an assay dependent concentration.|
|Small Molecule Fluorescent Optical Detection||Small Molecule Fluorescent Optical Detection: Use at an assay dependent concentration.|
|Receptor Localization||Receptor Localization: Use at an assay dependent concentration.|
NAN-190 Fluorescent ligand (Red) images
Left: ab118177 ligand (100 nM) binding to live CHO cells expressing serotonergic 5-HT1A receptors. Right: Binding blocked by the unlabelled competitor WAY-100635 (10 µM). Nuclei have been counter-stained with Hoechst.
To determine the antagonist activity of ab118177, serial dilutions of the agonist was added to wells with or without ab118177. The agonist Ca++ response in the absence or presence of ab118177 was determined by exciting at 340nm and 380 nm and ratioing the fluorescence intensity of Fura-2 signal collected at 320 nm. The apparent KD was calculated from the rightward shift of the agonist response curve in the presence of ab118177, compared to the response curve for the agonist alone. KD values Antagonist –log; KD values 5-HT1A 8.75: 5-HT2A 6.34
References for NAN-190 Fluorescent ligand (Red) (ab118177)
ab118177 has not yet been referenced specifically in any publications.