WB result of Histone H3 (di methyl K79) Rabbit pAb
Primary antibody: Histone H3 (di methyl K79) Rabbit pAb at 1/1000 dilution
Lane 1: HeLa whole cell lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 17 kDa
Observed MW: 17 kDa
Exposure time: 3 s (This blot was developed with high sensitivity substrate)
Histone H3 (di methyl K79) Rabbit polyclonal antibody
Histone H3 (di methyl K79) Rabbit polyclonal antibody
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Synonyms | H3K79me2 |
| Immunogen | Synthetic Peptide |
| Location | Nucleus |
| Accession | P68431 |
| Antibody Type | Polyclonal antibody |
| Isotype | IgG |
| Application | WB, ICC, ICFCM, ChIP |
| Reactivity | Hu, Ms |
| Predicted Reactivity | Fu, Pl, Ys, SeUr, Dr, C.el, Hm, Or, Bv, Xe |
| Purification | Protein A |
| Concentration | 0.8 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| Dot Blot | 1:1000 | |
| WB | 1:1000 | |
| ICC | 1:500 | |
| ICFCM | 1:800 | |
| ChIP | 1:20-1:50 |
Background
H3K79me2 is an epigenetic modification to the DNA packaging protein Histone H3. It is a mark that indicates the di-methylation at the 79th lysine residue of the histone H3 protein. H3K79me2 is detected in the transcribed regions of active genes.
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Western Blot
WB result of Histone H3 (di methyl K79) Rabbit pAb
Primary antibody: Histone H3 (di methyl K79) Rabbit pAb at 1/1000 dilution
Lane 1: Neuro-2a whole cell lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 17 kDa
Observed MW: 17 kDa
Exposure time: 3 s (This blot was developed with high sensitivity substrate)
FC
Flow cytometric analysis of 4% PFA fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) labelling Histone H3 (di methyl K79) antibody at 1/800 dilution (0.1 μg)/ (Red) compared with a Rabbit monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti - Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody.
Dot Blot
Dot blot result of Histone H3 (di methyl K79) Rabbit pAb
Lane 1: H3 (di methyl K79) peptide
Lane 2: H3 (mono methyl K79) peptide
Lane 3: H3 (tri methyl K79) peptide
Lane 4: H3 unmodified peptide
Primary antibody: Histone H3 (di methyl K79) Rabbit pAb at 1/1000 dilution
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Exposure time: 90 s
Immunocytochemistry
ICC shows positive staining in HeLa cells. Anti-Histone H3 (di methyl K79) antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
ChIP
Chromatin immunoprecipitation (ChIP) was
performed on HeLa cells cross - linked with
1% formaldehyde for 10 min, then chromatin
was fragmented by sonication.
Parallel reactions used Histone H3 (di methyl
K79) Recombinant Rabbit mAb (S-861-22) and
IgG Isotype Control (SDT-R173) at 1:50 for
immunoprecipitation.
Post - immunoprecipitation, both samples
were washed, eluted, and cross - links reversed.
Purified DNA was analyzed by qPCR.
qPCR showed the enrichment of RPL30, GAPDH,
MYOD1, AFM, SAT-α and SAT-2 in Histone H3
(di methyl K79) Recombinant Rabbit mAb (S-861-22)-
immunoprecipitated sample.
