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Our Abpromise guarantee covers the use of ab66219 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||1/100 - 1/400.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 41 kDa (predicted molecular weight: 41 kDa).Can be blocked with Nociceptin receptor peptide (ab129205).|
|IHC-Fr||1/40 - 1/100.|
Sk-N-SH cells were incubated at 37°C for 24h with vehicle control (0 µM) and 1 µM of kainic acid (ab120100). Decrease expression of nociceptin receptor in SK-N-SH cells correlates with an increase in kainic acid concentration, as described in literature.
Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 30µg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab66219 at 1 µg/ml and ab8227 at 1 µg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 dilution and visualised using ECL development solution.