U2OS cells were stained with the Bioss antibody against H3K36me1 (cat. No. bs-53131R) and with DAPI. Cells were fixed with 4% formaldehyde for 20’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure A: cells were immunofluorescently labeled with the H3K36me1 antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure B, C, D, and E: staining of the cells with the H3K36me1 antibody after incubation of the antibody with 2 ng/μl blocking peptide containing the unmodified and the mono-, di- and trimethylated H3K36, respectively.


Histone extracts (15 μg) from HeLa cells were analyzed by Western blot using the Bioss antibody against H3K36me1 (cat. No. bs-53131R) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk.


A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K36me1 (cat. No. bs-53131R) with peptides containing other modifications of histone H3 or the unmodified H3K36. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000.


To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody directed against H3K36me1 (cat. No. bs-53131R), crude serum and Flow Through. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:46,000.