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Synthetic phosphopeptide corresponding to amino acids surrounding phospho-286 from Rat brain CaMKII.
Our Abpromise guarantee covers the use of ab32678 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000. Predicted molecular weight: 50 kDa. Predicted molecular weight: ~50 kDa for the alpha subunit and ~60 kDa for the beta subunit of CaMKII.|
|IHC-P||Use a concentration of 2 µg/ml.|
|IHC-Fr||Use at an assay dependent concentration.|
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Blocked with 5% milk for 1 hour at RT.
Incubated with primary antibody for 14 hours at 4°C in TBS-T20.
PC 12 cells were incubated at 37°C for 30 minutes with vehicle control (0 µM) and different concentrations of myricetin (ab120721). Increased expression of CaMKII (phospho T286) (ab32678) in PC 12 cells correlates with an increase in myricetin concentration, as described in literature.
Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 20µg 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 3% milk before being incubated with ab32678at 1/500 dilution and ab52476 at 1/500 dilution 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.
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