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Anti-HIST3H3 Antibody Y47
Also for HIST3H3 (NM_003493)
|A synthetic (Dimethyl-K)-peptide corresponding to residues surrounding Lys4 of Histone H3 was used as immunogen. The antibody only detects Histone H3 dimethylated on Lysine 4. Predicted to cross-react with most species, based on sequence homology.|
||0.5~1.0 mg/ml (Lot Dependent)
|WB, IHC, IF, FC
||ChIP: Use at an assay dependent dilution; CHIPseq: Use at an assay dependent dilution; IHC-Fr: Use at an assay dependent concentration; WB: 1:1000 - 1:10000; IHC-P: Use at an assay dependent dilution; ICC/IF: 1:250; IP: Use at an assay dependent concentration; FC: 1:100
|PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%|
|Homo sapiens histone cluster 3, H3 (HIST3H3)|
|H3.4; H3/g; H3FT; H3t|
|Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. This gene is intronless and encodes a member of the histone H3 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element. This gene is located separately from the other H3 genes that are in the histone gene cluster on chromosome 6p22-p21.3. |
EGFR1 Signaling Pathway
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Western blot - Histone H3 (di methyl K4) antibody [Y47]; All lanes : Anti-Histone H3 (di methyl K4) [Y47] antibody - ChIP Grade at 1/20000 dilution.Lane 1 : HeLa cell lysate.Lane 2 : Non methylated recombinant Histone H3.Predicted band size : 15 kDa.Observed band size : 17 kDa .
Immunohistochemistry (Paraffin-embedded sections) - Histone H3 (di methyl K4) antibody [Y47]; Ab32356, at a 1/100 dilution, staining Histone H3 in paraffin embedded breast carcinoma tissue sections by Immunohistochemistry.
Immunocytochemistry/ Immunofluorescence - Histone H3 (di methyl K4) antibody [Y47]; Ab32356, at a 1/250 dilution, staining Histone H3 in HeLa cells by Immunofluorescence.
Immunocytochemistry/ Immunofluorescence - Histone H3 (di methyl K4) antibody [Y47] - ChIP Grade; ab32356 (1/200) staining Histone H3 (di methyl K4) in HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilized with 0.5% Triton X100 and counterstained with DAPI in order to highlight the nucleus. For further experimental details please refer to Abreview.
Immunocytochemistry/ Immunofluorescence - Histone H3 (di methyl K4) antibody [Y47] - ChIP Grade; ICC/IF image of ab32356 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody overnight at +4°C. The secondary antibody (green) was DyLight? 488 goat anti-rabbit IgG - H&L, pre-adsorbed used at a 1/250 dilution for 1h. Alexa Fluor? 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Flow Cytometry-Anti-Histone H3 (di methyl K4) antibody [Y47] - ChIP Grade(ab32356); Overlay histogram showing HeLa cells stained with ab32356 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody for 30 min at 22°C. The secondary antibody used was DyLight? 488 goat anti-rabbit IgG (H+L) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x10^6 cells) used under the same conditions. Acquisition of >5,000 events was performed.
ChIP - Histone H3 (di methyl K4) antibody [Y47]; Chromatin was prepared from Hela cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25µg of chromatin, 8µl of ab32356 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach). Primers and probes are located in the first kb of the transcribed region. .