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Immunogen was a synthetic peptide, which represented a portion of human E1A binding protein p300 encoded within exons 15 and 16 (LocusLink ID 2033).
Our Abpromise guarantee covers the use of ab10485 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||Use at 1-4 µg/mg of lysate.|
|WB||1/5000 - 1/25000. Can be blocked with KAT3B / p300 peptide (ab4914).|
|IHC-P||1/1000 - 1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|CHIPseq||Use at an assay dependent concentration.|
Left-hand panel represents a coomassie-blue stained gel loaded with 10mg of HeLa cytosolic (S100) and nuclear (NE) extracts;
For the Immunoprecipitation experiment (right-hand panel), ab10485 was used at a concentration of 20 µg/10 mg of HeLa whole cell lysate (lanes correspond to 5, 15 and 50 µg of lysate). The flow-through was analysed by WB using ab10485 at a concentration of 0.04 µg/ml.
The identity of the band marked as "p300" was also confirmed using mass spectrometry.
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