Anti-Histone H3 (di methyl K4) antibody - ChIP Grade

8 product images

• 

  Peptide Array - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)

  All batches of ab7766 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 - di methyl K4 peptide (Human Histone H3 (di methyl K4) peptide ab7768), indicating that this antibody specifically recognises the Histone H3 - di methyl K4 modification.
  

  
  
  1. Human Histone H3 (mono methyl K4) peptide ab1340 - Histone H3 - mono methyl K4
  
  
  2. Human Histone H3 (tri methyl K4) peptide ab1342 - Histone H3 - tri methyl K4
  
  
  3. Human Histone H3 (mono methyl K9) peptide ab1771 - Histone H3 - mono methyl K9
  
  
  4. Human Histone H3 (di methyl K9) peptide ab1772 - Histone H3 - di methyl K9
  
  
  5. Human Histone H3 (tri methyl K9) peptide ab1773 - Histone H3 - tri methyl K9
  
  
  6. Human Histone H3 (mono methyl K27) peptide ab1780 - Histone H3 - mono methyl K27
  
  
  7. Human Histone H3 (di methyl K27) peptide ab1781 - Histone H3 - di methyl K27
  
  
  8. Human Histone H3 (tri methyl K27) peptide ab1782 - Histone H3 - tri methyl K27
  
  
  9. Human Histone H3 (unmodified) peptide ab7228 - Histone H3 - unmodified
  
  
  10. Human Histone H3 (di methyl K4) peptide ab7768 - Histone H3 - di methyl K4
  
  
• 

  Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)

  ab7766 staining Histone H3 (di methyl K4) in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab7766 at 5µg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

  Also suitable in cells fixed with 100% methanol (5 min).

  Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• 

  This image is courtesy of Kirk McManus in the lab of Michael Hendzel, Univeristy of Alberta

  Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)

  Dimethylated lysine 4 (green) is found in several hundred small nuclear foci that do not colocalize with condensed regions of chromatin (DAPI stained, red). The perinuclear and perinucleolar heterochromatin do not stain with this antibody.

• 

  Western blot - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)

  All lanes: Western blot - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766) at 1 µg/mL

  All lanes: Calf Thymus Histone Preparation Nuclear Lysate (ab121) at 0.5 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (Goat Anti-Rabbit IgG H&L (HRP) preadsorbed ab97080) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 15 kDa

  Observed band size: 14 kDa, 17 kDa

  Exposure time: 4min

• 

  This image is courtesy of a customer review submitted by Dr Alexander Rapp

  Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)

  ab7766 at 1/200 staining human U2OS (osteosarcoma) cells by ICC/IF. The cells were paraformaldehyde fixed and then stained with the antibody for 1 hour. A Cy2 ® conjugated donkey anti-rabbit antibody was used as the secondary (green). The image shows uniformal staining of the whole nucleus, with several specles found. The insert shows H3-di methyl K4 of tw2 cells only. DAPI nuclear staining is shown in blue.

• 

  Western blot - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)

  Blocking buffer: 2% BSA
  

  Gel type: MES

  Exposure Time: 1 minute

  All lanes: Western blot - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766) at 1 µg/mL

  Lane 1: HeLa nuclear lysate (triton enriched) at 20 µg

  Lane 2: NIH 3T3 nuclear lysate (triton enriched) at 20 µg

  Lane 3: PC12 nuclear lysate (triton enriched at 20 µg

  Secondary

  All lanes: Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution

  Predicted band size: 15 kDa

  Observed band size: 17 kDa

• 

  Functional Studies - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)

  MCF7 cells were incubated at 37°C for 24h with vehicle control (0 μM) and different concentrations of tranylcypromine hydrochloride (Tranylcypromine hydrochloride (2-PCPA), Irreversible monoamine oxidase (MAO) inhibitor ab120606). Increased expression of Histone 3 K4 di-methyl (ab7766) in MCF7 cells correlates with an increase in tranylcypromine hydrochloride concentration, as described in literature.
  

  Nuclear extracts were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10 μg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab7766 at 1 μg /ml and Anti-Histone H3 antibody - Nuclear Marker and ChIP Grade ab1791 at 1 μg /ml overnight at 4°C. Antibody binding was detected using an anti-rabbit HRP secondary antibody (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/10000 dilution and visualized using ECL development solution.

• 

  ChIP - Anti-Histone H3 (di methyl K4) antibody - ChIP Grade (ab7766)

  Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25μg of chromatin, 2μg of ab7766 (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 for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.