5-fC | 5-formylcytosine
AS16 3165 | clonality: polyclonal | host: rabbit | reactivity:human, mouse, other species
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|Recommended dilution||3 ĩl (DIP), 1 : 10 000 (ELISA)|
|Expected | apparent MW|
|Predicted reactivity||Mouse, broad species range|
|Not reactive in||No confirmed exceptions from predicted reactivity are currently known.|
|Selected references||To be added when available, antibody released in February 2016.|
DIP: HEK293 cells were transfected with a reporter gene and hydroxymethylated in vitro with either a pCAG expression vector containing the TET2 catalytic domain (TET2cd) or a negative control pCAG vector. DIP assays were performed on 4 μg of sheared and denatured DNA using 3 μl of the polyclonal antibody against 5-fC in a total of 500 μl IP buffer. QPCR was performed with primers specific for the reporter gene. Figure 1 shows the recovery, expressed as a % of input (mean +standard deviation of 3 different experiments).
ELISA: to determine the titer of the antibody, an ELISA was performed using a serial dilution of the polyclonal antibody directed against 5-fC. The plates were coated with the immunogen. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be >1:100 000.
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