Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - BSA and Azide free

10 product images

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  ChIP - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - BSA and Azide free (ab222481)

  This ChIP data was generated using the same anti-Histone H3K27me3 antibody clone, EPR18607, in a different buffer formulation (cat# Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade ab192985).
  

  Chromatin was prepared from HeLa (Human epithelial cell line from cervix adenocarcinoma) 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 Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade ab192985 (blue), and 20μl of Anti rabbit IgG sepharose beads. 2μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

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  Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - BSA and Azide free (ab222481)

  This ICC/IF data was generated using the same anti-Histone H3K27me3 antibody clone, EPR18607, in a different buffer formulation (cat# Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade ab192985).
  

  Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Histone H3 (tri methyl K27) with Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade ab192985 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counterstain is DAPI (blue).

  Tubulin is detected with Anti-alpha Tubulin mouse MAb (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) secondary antibody at 1/1000 dilution (red).

  The negative controls are as follows:-
  -ve control 1: Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade ab192985 at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) secondary antibody at 1/1000 dilution.
  -ve control 2: Anti-alpha Tubulin mouse MAb (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - BSA and Azide free (ab222481)

  Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Histone H3 (tri methyl K27) with Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade ab192985 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Nuclear staining on human colon tissue is observed. Counter stained with Hematoxylin.
  

  Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade ab192985).

  Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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  Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - BSA and Azide free (ab222481)

  Clone EPR18607 (ab222481) has been successfully conjugated by Abcam. This image was generated using Anti-Histone H3 (tri methyl K27) antibody [EPR18607] (Alexa Fluor® 647). Please refer to Alexa Fluor® 647 Anti-Histone H3 (tri methyl K27) antibody [EPR18607] ab270163 for protocol details.

  Immunofluorescence staining of Histone H3 (tri methyl K27) in HeLa cells using Alexa Fluor® 647 Anti-Histone H3 (tri methyl K27) antibody [EPR18607] ab270163. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% 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 Alexa Fluor® 647 Anti-Histone H3 (tri methyl K27) antibody [EPR18607] ab270163 at 1/100 dilution (shown in red) and Alexa Fluor® 488 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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  Peptide Array - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - BSA and Azide free (ab222481)

  Peptide array analysis was performed using Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade ab192985 at a concentration of 0.1ug/ml, followed by Goat Anti-Rabbit IgG, (H+L), Fluo 647nm conjugated secondary antibody at a 1/50000 dilution.

  Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade ab192985 was tested in Peptide Array against 501 different modified and unmodified histone peptides; each peptide is printed on the array at six concentrations (each in triplicate).

  Circle area represents affinity between the antibody and a peptide: all antigen-containing peptides are displayed as red circles, all other peptides as blue circles. The affinity is calculated as area under curve when antibody binding values are plotted against the corresponding peptide concentration. Each circle area is normalized to the peptide with the strongest affinity.

  The complete dataset, including full list of all peptides and information on the position of each peptide in the diagram, is available under the Support & downloads section.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade ab192985).

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  ELISA - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - BSA and Azide free (ab222481)

  ELISA analysis was performed on 1ug/ml of antigen using Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade ab192985 at a concentration range of 0-0.25ug/ml, followed by Alkaline Phosphatase-conjugates AffiniPure Goat anti-rabbit IgG (H&L) secondary antibody at a 1/2500 dilution.

  All batches of Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade ab192985 are tested in ELISA against peptides to different Histone H3 modifications. Results show strong binding to Histone H3 - tri methyl K27 immunizing peptide, indicating that this antibody specifically recognizes the Histone H3 - tri methyl K27 modification. Weak binding (14%) was also detected against H3 - di methyl K27 modification.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade ab192985).

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  Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - BSA and Azide free (ab222481)

  Clone EPR18607 (ab222481) has been successfully conjugated by Abcam. This image was generated using Anti-Histone H3 (tri methyl K27) antibody [EPR18607] (Alexa Fluor® 488). Please refer to Alexa Fluor® 488 Anti-Histone H3 (tri methyl K27) antibody [EPR18607] ab270162 for protocol details.

  Immunofluorescence staining of Histone H3 (tri methyl K27) in HeLa cells using Alexa Fluor® 488 Anti-Histone H3 (tri methyl K27) antibody [EPR18607] ab270162. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% 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 Alexa Fluor® 488 Anti-Histone H3 (tri methyl K27) antibody [EPR18607] ab270162 at 1/100 dilution (shown in green) and Alexa Fluor® 647 Anti-Tubulin antibody [YOL1/34] - Microtubule Marker ab195884, Rat monoclonal to Tubulin (Alexa Fluor^® 647), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - BSA and Azide free (ab222481)

  Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling Histone H3 (tri methyl K27) with Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade ab192985 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Nuclear staining on mouse colon tissue is observed. Counter stained with Hematoxylin.
  

  Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade ab192985).

  Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - BSA and Azide free (ab222481)

  Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling Histone H3 (tri methyl K27) with Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade ab192985 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Nuclear staining on rat kidney tissue is observed. Counter stained with Hematoxylin.
  

  Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade ab192985).

  Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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  ChIP-sequencing - Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - BSA and Azide free (ab222481)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade ab192985).
  Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 107 HeLa cells and 4 µg of Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade (Anti-Histone H3 (tri methyl K27) antibody [EPR18607] - ChIP Grade ab192985). ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. Additional screenshots of mapped reads can be downloaded here.