The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration. PubMed: 19000314
Use a concentration of 1 µg/ml.
1/1000. Predicted molecular weight: 344 kDa.
Component of the NuA4 histone acetyltransferase complex which is involved in transcriptional activation of select genes principally by acetylation of nucleosomal histones H4 and H2A. This modification may both alter nucleosome - DNA interactions and promote interaction of the modified histones with other proteins which positively regulate transcription. May be required for transcriptional activation of E2F1 and MYC target genes during cellular proliferation. The NuA4 complex ATPase and helicase activities seem to be, at least in part, contributed by the association of RUVBL1 and RUVBL2 with EP400. May regulate ZNF42 transcription activity.
ICC/IF image of ab5201 stained human HeLa cells. The cells were PFA fixed (3.7% PFA, 10 min) and incubated with the antibody (ab5201, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
Western blot - Anti-p400 antibody (ab5201)
All lanes : Anti-p400 antibody (ab5201)
Lane 1 : Mock Transfected HeLa Lane 2 : Tip60-TAP Transfected HeLa Lane 3 : ING4-TAP Transfected HeLa
Predicted band size: 344 kDa
Rabbit polyclonal to p400 used at 1/1000. p400 is part of the Tip60/NuA4 HAT complex.
Nuclear extracts from Mock (1), Tip60 (2) and an unrelated TAP-tagged protein (3) were purified by tandem affinity purification. No non-specific bands are observed.
Whilst it is our normal policy to display the full gel for Western Blots, in this case we received a strip image from an academic collaborator. However, we still feel this image provides useful information on ab5201.
Image from PMID:16387653
Western blot analysis of p400 antibody using TAP purifications of ING4 and Tip60 that shows two distinct complexes