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Our Abpromise guarantee covers the use of ab12169 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ChIP||Use at an assay dependent concentration. PubMed: 24919190|
|IHC-Fr||1/1000. PubMed: 20407577|
|IHC-P||Use at an assay dependent concentration.|
|ELISA||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration.|
|WB||Use a concentration of 0.25 - 0.5 µg/ml. Detects a band of approximately 55 kDa.|
Chromatin was isolated and pooled from the two PF cortices of a single mouse. ChIP experiments were performed using a minimum of 3 mice per group. Chromatin was sheared in an ice bath by a 25 cycles of 30 sec on/off sonication using the “Bioruptor UCD-20” sonicator. Samples were kept on ice during 30 s between two pulses. An aliquot of precleared chromatin was collected as the input. The samples were incubated overnight at 4°C with 5 µl ab12169. The immunoprecipitated chromatin was analyzed by quantitative PCR with primers designed to amplify short regions of the promoters of genes of interest.
ChIP assay on c-Fos gene promoter using ab12169 antibody. Results show a significant lower enrichment of HDAC 2 in PF cortex of (amyloid precursor protein) APP−/− mouse, Student’s t-test: **p<0.01. IgG was used as negative control. Equal amounts of ChIP and input chromatin were used for qRT-PCR analysis on the c-Fos gene promoter. Enrichment values were normalized to input values and represented the average of three or more mice per experiment. Results are expressed as mean ± SD.
Western Blot using Rabbit anti-HDAC2
This image is courtesy of an anonymous abreview.