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Epigenetics

Epigenetics

Our Epigenetics Research Area is designed to enable investigation of transcriptional control, chromatin remodeling, histone modification patterns, and DNA methylation dynamics across diverse experimental workflows. This collection includes antibodies for use in WB, IHC, ICC, IF, FC, ELISA, and other common applications.



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Human osteosarcoma (U2OS) cells were stained with the Bioss antibody against H3K79me3 (Cat. No. bs-53121R) and with DAPI. Cells were fixed with 2.5% formaldehyde for 30\u2019 and blocked with PBS\/TX-100 containing 1% BSA. Figure A: cells were immunofluorescently labeled with the H3K79me3 antibody (left) diluted 1:300 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right). Figure B and C: staining of the cells with the H3K79me3 antibody after incubation of the antibody with 2 ng\/\u03bcl H4K20me3 and H3K79me3 peptide, respectively.
Histone extracts of HeLa cells (15 \u03bcg) were analyzed by Western blot using the Bioss antibody against H3K79me3 (Cat. No. bs-53121R), diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The molecular weight marker is shown on the right; the location of the protein of interest is indicated on the left.
A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K79me3 (Cat. No. bs-53121R) with peptides containing other histone modifications and the unmodified H3K79. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000.The figure shows a high specificity of the antibody for the modification of interest.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of Bioss antibody directed against H3K79me3 (Cat. No. bs-53121R), crude serum and flow through 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 1:3,500.

Catalog #1: bs-53121R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human, Mouse, Yeast

Conjugate: Unconjugated

Western blot was performed on whole cell (25 \u03bcg, lane 1) and histone extracts (15 \u03bcg, lane 2) from HeLa cells, and on 1 \u03bcg of recombinant histone H3 (lane 3) using H3K79me1 Polyclonal Antibody (Cat. No. bs-53126R). The antibody was diluted 1:200 in TBS-Tween containing 5% skimmed milk.
A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K79me1 (Cat. No. bs-53126R) with peptides containing other histone modifications and the unmodified H3K79. One hundred to 0.2 pmol of the respective peptides 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 H3K79me1 (Cat. No. bs-53126R). 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 1:11,500.
ChIP assays were performed using human HeLa cells, the Bioss antibody against H3K79me1 (Cat. No. bs-53126R) and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 1 million cells. A titration consisting of 1, 2, 5 and 10 \u03bcg of antibody per ChIP experiment was analyzed. IgG (2 \u03bcg\/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for a region 1 kb upstream of the promoter and the coding region of the active GAPDH gene, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene and the Sat2 satellite repeat, used as negative controls. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Catalog #1: bs-53126R

Applications: WB, ELISA, dot-blot, ChIP-seq

Reactivity: Human, Yeast

Conjugate: Unconjugated