The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
ab17722 staining FMRP in SK-N-SH cells treated with (R,S)-3,5-DHPG (ab120020), by ICC/IF. Increase in FMRP expression correlates with increased concentration of (R,S)-3,5-DHPG, as described in literature. The cells were incubated at 37°C for 1h in media containing different concentrations of ab120020 ((R,S)-3,5-DHPG) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab17722 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
References for (R,S)-3,5-DHPG (ab120020)
This product has been referenced in:
Lin FF et al. Cloning and stable expression of the mGluR1b subtype of human metabotropic receptors and pharmacological comparison with the mGluR5a subtype. Neuropharmacology36:917-31 (1997).
Read more (PubMed: 9257936) »
Brabet I et al. Phenylglycine derivatives discriminate between mGluR1- and mGluR5-mediated responses. Neuropharmacology34:895-903 (1995).
Read more (PubMed: 8532171) »