Anti-Histone H3 (asymmetric di methyl R17) antibody - ChIP Grade

• Supplier: Abcam
• Catalog: ab8284
• Prices: $403.00
• Sizes: 50 µg
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Applications: ChIP ChIP Array IHC-P ICC/IF ICC/IF ChIP DB IP WB
• Species Reactivities: Mouse, Rat, Chicken, Bovine, Human, C. elegans, Drosophila, Toxoplasma gondii, Mammalian
• Antigen: Synthetic peptide corresponding to Human Histone H3 aa 1-100 (asymmetric di methyl R17) conjugated to Keyhole Limpet Haemocyanin (KLH)
• Blocking Peptide: Human Histone H3 (asymmetric di methyl R17) peptide
• Description: Rabbit Polyclonal
• Gene: HIST3H3
• Conjugate: Unconjugated

Product images

All batches of ab8284 are tested in Peptide Array against peptides to different Histone H3 modifications. Six dilutions of each peptide are printed on to the Peptide Array in triplicate and results are averaged before being plotted on to a graph. Results show strong binding to Histone H3 - asymmetric di methyl R17 peptide (ab16935), indicating that this antibody specifically recognises the Histone H3 - asymmetric di methyl R17 modification.ab16935 - Histone H3 - asymmetric di methyl R17ab32948 - Histone H3 - symmetric di methyl R17ab14663 - Histone H3 - unmodified

Anti-Histone H3 (asymmetric di methyl R17) antibody - ChIP Grade (ab8284) at 1 µg/ml + HeLa Histone Preparation Nuclear Lysate at 2.5 µg/mlSecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.

Total U2OS cell extract was western blotted using the anti-Me-R17H3 antibody. The asterisk indicates methylated histone H3. The left panel shows presence of core histones (indicated on the left) by Coomassie Blue staining. Molecular weights are indicated on the right.

A dot blot was performed using unmodified peptide (lane 1), Histone H3 mono methyl R17 peptide (lane 2), Histone H3 asymmetric di methyl R17 peptide (lane 3) and Histone H3 symmetric di methyl R17 peptide (lane 4). The dot blot indicates that ab8284 is specific to Histone H3 asymmetric di methyl R17.

ab8284 was used in immunohistochemistry with paraffin embedded sections of human tonsil, using DAB as a chromogen (brown). Counterstaining of nuclei was performed with haemotoxylin (blue).Staining is seen confined to the nucleus.

ICC/IF image of ab8284 stained MCF7 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 (ab8284, 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.

Product References

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