H3K79me2 Polyclonal Antibody
Catalogue Number: BS-53111R-BSS
BS-53111R Image
BS-53111R Image
BS-53111R Image
BS-53111R Image
BS-53111R Image
BS-53111R Image
ChIP assays were performed using HeLa cells, the Bioss antibody against H3K79me2 (Cat. No. bs-53111R) and optimized PCR primer pairs for qPCR. ChIP was performed with an Auto Histone ChIP-seq kit, using sheared chromatin from 1 million cells. A titration consisting of 0.5, 1, 2 and 5 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the coding regions of the active EIF2S3 and CCT5 genes, used as positive controls, and for the inactive MYOD1) 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).
BS-53111R Image
ChIP was performed with 1 µg of the Bioss antibody against H3K79me2 (Cat. No. bs-53111R) as described above. The IP'd DNA was subsequently analyzed on an Illumina HiSeq 2000. Library preparation, cluster generation, and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. The figure shows the peak distribution along the complete sequence and a 5 Mb region of chromosome 1 (figure A and B) and in two 300 kb regions surrounding the EIF2S3 and CCT5 positive control genes (figure C and D).
BS-53111R Image
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody directed against H3K79me2 (Cat. No. bs-53111R). 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 1:6,600.
BS-53111R Image
A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K79me2 (Cat. No. bs-53111R) with peptides containing other modifications and unmodified sequences of histone H3. 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 1:5,000. The figure shows a high specificity of the antibody for the modification of interest.
BS-53111R Image
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 H3K79me2 (Cat. No. bs-53111R) diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.
| Manufacturer: | Bioss Inc |
| Shelf Life: | 12 months |
| Preservative: | 0.05% Sodium azide; ProClin™ 300 |
| Type: | ChIP Validated Antibodies |
| Shipping Condition: | Blue Ice |
| Unit(s): | 50 ug |
| Host name: | Rabbit |
| Clone: | |
| Isotype: | |
| Immunogen: | Polyclonal antibody raised in rabbit against the region of histone H3 containing the dimethylated lysine 79 (H3K79me2), using a KLH-conjugated synthetic peptide |
| Application: | ELISA, ChIP, WB, DB |
Description
Description: Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.
Additional Text
Uniprot ID
P68431
Purification
Affinity Purified
Gene ID
8350; 8351; 8352; 8353; 8354; 8355; 8356; 8357; 8358; 8968
Antibody Clonality
Polyclonal
Concentration
1.75 ug/ul
Modification
Unmodified
Gene Name
H3C1, H3C10, H3C11, H3C12, H3C2, H3C3, H3C4, H3C6, H3C7, H3C8
Storage Note
Store at -20°C for 12 months.
bs-53111R Datasheet
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