Our Abpromise guarantee covers the use of ab2831 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 2 µg/ml. Predicted molecular weight: 155 kDa.Can be blocked with SRC3 peptide (ab4915). Can be blocked with AIB1 peptide or reuse.|
|IHC-P||Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|ICC/IF||Use at an assay dependent concentration.|
|ChIP||Use 3µg for 106 cells. PubMed: 18499756|
ab2831 can be used in ChIP. You can expect recruitment of AIB1on the pS2 promoter at some point during transcriptional activation. The amount of DNA precipiated is significant.
Sonicated Chromatin prepared from untreated or 17beta-estradiol (E2) treated MCF7 cells was subjected to the ChIP procedure with ab2831 to AIB1 and the immunoprecipitated chromatin was analysed in the proximal region of the estrogen-responsive pS2 promoter (as shown above) and quantified by real-time PCR (values are % of inputs). The primers are designed to follow the nucleosome E (including the Estrogen Responsive Element ERE). 3
ICC/IF image of ab2831 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab2831, 5µg/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).