5-hmC Antibody (C.15200200) - BSA Free

Dot Blot: 5-hmC Antibody (C.15200200) [NBP2-59177] - 5-Hydroxymethylcytosine Antibody (C.15200200) [NBP2-59177] - Dot Blot analysis of the antibody with the C, mC and hmC PCR controls 200 to 2 ng (equivalent of 10 to 0.1 pmol of C-bases) of the hmC (1), mC (2) and C (3) PCR controls from the antibody (dilution 1:500). The membranes were exposed for 30 seconds.

Methylated DNA Immunoprecipitation: 5-hmC Antibody (C.15200200) [NBP2-59177]

Methylated DNA Immunoprecipitation: 5-hmC Antibody (C.15200200) [NBP2-59177] - 5-Hydroxymethylcytosine Antibody (C.15200200) [NBP2-59177] - An hydroxymethylated DNA IP (hMeDIP) was performed using the antibody directed against 5-hydroxymethylcytosine. The IgG isotype antibodies from mouse was used as negative control. The DNA was prepared with the GenDNA module of the hMeDIP kit and sonicated to have DNA fragments of 300-500 bp. 1 ug of human HeLa cells DNA were spiked with non-methylated, methylated, and hydroxymethylated PCR fragments. The IP'd material has been analysed by qPCR using the primer pair specific for the 3 different control sequences. The obtained results show that the mouse monoclonal for 5-hmC is highly specific for this base modification (no IP with non-methylated or methylated C bases containing fragments).

ELISA: 5-hmC Antibody (C.15200200) [NBP2-59177] - To determine the titer, an ELISA was performed using a serial dilution of the antibody directed against 5-hmC in antigen coated wells. The antigen used was KHL coupled to 5-hmC base. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:40,000.