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Our Abpromise guarantee covers the use of ab28450 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use at an assay dependent concentration.|
|WB||Use a concentration of 0.5 µg/ml. Predicted molecular weight: 212 kDa. Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.|
|ChIP||Use 5-10 µg for 25 µg of chromatin.|
Quiescent human colon carcinoma HCT116 cultures were treated with 10% FBS for three time points (0, 15, 30 minutes) or (0, 30, 60 minutes) were used in Matrix-ChIP and real-time PCR assays at EGR1 gene (Exon1) and 15kb upstream site.
Chromatin was prepared from Hela cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10mins. The ChIP was performed with 25µg of chromatin, 8µg of ab28450 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman and sybr green approach). Primers and probes for the actively transcribed genes GAPDH and ACT1 are located in the core promoter.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"