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

5 product images

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K36) antibody - ChIP Grade (ab9050)

  ab9050 stained in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.
  

  Cells were fixed with 100% methanol for 5 minutes at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1 hour at room temperature to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab9050 at 0.1 μg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) (pseudo-colored red) and Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) secondary antibody (colored green) used at 1 μg/ml for 1 hour at room temperature.

  DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 μM for 1 hour at room temperature.

• 

  Western blot - Anti-Histone H3 (tri methyl K36) antibody - ChIP Grade (ab9050)

  All lanes: Western blot - Anti-Histone H3 (tri methyl K36) antibody - ChIP Grade (ab9050) at 1 µg/mL

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

  Secondary

  All lanes: Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

  Performed under reducing conditions.

  Predicted band size: 15 kDa

  Observed band size: 17 kDa

• 

  Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K36) antibody - ChIP Grade (ab9050)

  ab9050 staining Histone H3 (tri-methyl K36) in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.
  

  The cells were fixed with 100% methanol for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1 hour. The cells were then incubated with ab9050 at 0.1 μg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti beta-Tubulin) at 1 μg/ml overnight at +4°C, followed by a further incubation at room temperature for 1 hour with a goat anti-rabbit AlexaFluor^®488 secondary (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat anti-mouse AlexaFluor^®594 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labeled in blue with DAPI.

  Negative controls: 1– Rabbit primary antibody and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.

• 

  This image is courtesy of an anonymous customer review

  Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K36) antibody - ChIP Grade (ab9050)

  ab9050 staining Histone H3 (tri methyl K36) in Saos-2 (Human osteosarcoma cell line) cells by ICC/IF (Immunocytochemistry/immunofluorescence).
  

  Cells were fixed with paraformaldehyde, permeabilized with 0.25% Triton in PBS and blocked with 1% BSA for 1 hour at room temperature. Samples were incubated with primary antibody (1/1000) for 1 hour. An Alexa Fluor^®488-conjugated Goat anti-rabbit IgG polyclonal (1/250) was used as the secondary antibody.

• 

  ChIP - Anti-Histone H3 (tri methyl K36) antibody - ChIP Grade (ab9050)

  Chromatin was prepared from U-2 OS (Human bone osteosarcoma epithelial cell line) cells according to the Abcam X-ChIP protocol.
  

  Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25 μg of chromatin, 2 μg of ab9050 (blue), and 20 μl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified on the GAPDH (active) and MYO-D (inactive) promoters and over the γ-Actin gene (active).

  Schematic diagram of the γ-Actin gene is shown on the top of the figure.

  Black boxes represent exons and thin lines represent introns.

  PCR products are depicted as bars under the gene.