Anti-Histone H3 (mono methyl K36) antibody - ChIP Grade

• Supplier: Abcam
• Catalog: ab9048
• Prices: $403.00
• Sizes: 100 µg
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Applications: IHC-P ICC/IF ICC/IF ICC/IF WB ChIP
• Species Reactivities: Mouse, Bovine, Human, Pig, Yeast, Arabidopsis thaliana, Drosophila, Mammalian
• Antigen: Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H3, mono methylated at K36
• Blocking Peptide: Human Histone H3 (mono methyl K36) peptide
• Description: Rabbit Polyclonal
• Gene: HIST3H3
• Conjugate: Unconjugated

Product images

ab9048 staining Histone H3 (mono methyl K36) in human differentiated haematopoietic stem cells by Flow Cytometry. Cells were fixed with paraformaldehyde and permeabilized with permeabilization buffer. The sample was incubated with the primary antibody (1/500) for 12 hours at 4°C. An Alexa Fluor® 488-conjugated goat polyclonal anti-rabbit IgG (1/1000) was used as the secondary antibody.Gating Strategy: Isotype negative control (white).See Abreview

All lanes : Anti-Histone H3 (mono methyl K36) antibody - ChIP Grade (ab9048) at 1/500 dilutionLane 1 : Histone prepLane 2 : Histone prep with Human Histone H3 (mono methyl K36) peptide (ab1783) at 1 µg/mlLane 3 : Histone prep with Human Histone H3 (di methyl K36) peptide (ab1784) at 1 µg/mlLane 4 : Histone prep with Human Histone H3 (tri methyl K36) peptide (ab1785) at 1 µg/mlLane 5 : Histone prep with Human Histone H3 (unmodified ) peptide (ab2623) at 1 µg/mlLane 6 : Histone prep with Human Histone H3 (mono methyl K4) peptide (ab1340) at 1 µg/mlLysates/proteins at 0.5 µg per lane.SecondaryGoat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilutionSecondary ab: Alexa Fluor 680 Goat anti-rabbit IgG 0.5µg histone prep used per lanePrimary antibody: Lane 1: ab9048 (Histone H3 Mono Methyl K36) 1/500Lane 2: ab9048 (Histone H3 Mono Methyl K36) 1/500 + ab1783 (ab9048) (Histone H3 Mono Methyl K36) peptide 1µg/ml Lane 3: ab9048 (Histone H3 Mono Methyl K36) 1/500 + ab1794 (ab9049) (Histone H3 Di

Staining of interphase nuclei of Hela cells with ab9048 (green) at a working dilution of 1/500. The DNA is stained with DAPI. ab9048 appears to be more associated with heterochromatin (DAPI intense regions) than euchromatin (DAPI less intense regions).

Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 min. The  ChIP was performed with 25 µg of chromatin, 2 µg of  ab9048 (blue), and 20 µl 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.

ab9048 staining Histone H3 (mono methyl K36) in Mouse pancreatic tumor tissue by Immunohistochemistry (Formalin/ PFA-fixed paraffin-embedded tissue sections). The sections were PFA-fixed and subjected to Heat mediated antigen retrieval in citrate and permeabilized with PBST prior to blocking with 5% serum for 1 hour at room temperature. The primary antibody was diluted 1/500 in PBS and incubated with the sample for 8 hours at 4°C. A Biotin-conjugated Goat anti-Rabbit polyclonal was used as the secondary antibody, diluted 1/1000.See Abreview

ICC/IF image of ab9048 stained HeLa cells. The cells were 4% formaldehyde fixed (10 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 (ab9048, 0.1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) 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. This antibody also gave a positive result in 4% formaldehyde fixed (10 min) Hek293, HepG2 and MCF7 cells at 0.1µg/ml, and in 100% methanol fixed (5 min) HeLa, Hek293, HepG2 and MCF7 cells at 0.1µg/ml.

All lanes : Anti-Histone H3 (mono methyl K36) antibody - ChIP Grade (ab9048) at 1/1400 dilutionLane 1 : Whole cell lysate prepared from Drosophila BG3 cellsLane 2 : Whole cell lysate prepared from Drosophila BG3 cellsLysates/proteins at 500000 cells per lane.SecondaryHRP donkey anti-rabbit monoclonal at 1/20000 dilutiondeveloped using the ECL techniqueObserved band size : 17,42 kDa (why is the actual band size different from the predicted?)Exposure time : 30 secondsImage courtesy of an anonymous Abreview.See Abreview

Product References

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