ChIP was performed as described above with 1 μg of the Bioss antibody against H3K4me1 (Cat. No. bs-53106R). The IP’d DNA was subsequently analyzed on an Illumina Genome Analyzer. Library preparation, cluster generation, and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure A and B show the enrichment in chromosomal regions surrounding the ACTB and GAS2L1 positive control genes. The position of the amplicon used in the qPCR validation is indicated by an arrow. Figure C and D show the H3K4me1 signal in two 1 Mb regions of chromosome 5, and X respectively.


To determine the titer, an ELISA was performed using a serial dilution of the Bioss antibody directed against H3K4me1 (Cat. No. bs-53106R) in antigen-coated wells. 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 1:20,100.


A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K4me1 (Cat. No. bs-53106R) with peptides containing other modifications or unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:10,000. The figure shows a high specificity of the antibody for the modification of interest.


Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using H3K4me1 Polyclonal Antibody (Cat. No. bs-53106R). The antibody was diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is shown on the right, the marker (in kDa) is shown on the left.


Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Bioss antibody against H3K4me1 (Cat. No. bs-53106R). The antibody was diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is shown on the right, the marker (in kDa) is shown on the left.


HeLa cells were stained with the Bioss antibody against H3K4me1 (cat. bs-53106R) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H3K4me1 antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.