Anti-Histone H3 (di methyl K36) antibody [EPR28335-73]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K36) antibody [EPR28335-73] (AB318964)

Immunohistochemical analysis of paraffin-embedded Human placenta tissue labeling Histone H3 (di methyl K36) with ab318964 at 1/2000 (0.267 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human placenta. The section was incubated with ab318964 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K36) antibody [EPR28335-73] (AB318964)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling Histone H3 (di methyl K36) with ab318964 at 1/500 (1.068 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing nuclear staining in HeLa cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Histone H3 (di methyl K36) antibody [EPR28335-73] (AB318964)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling Histone H3 (di methyl K36) with ab318964 at 1/500 dilution (0.1 ug)/Red compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K36) antibody [EPR28335-73] (AB318964)

Immunohistochemical analysis of paraffin-embedded Human prostatic hyperplasia tissue labeling Histone H3 (di methyl K36) with ab318964 at 1/2000 (0.267 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human prostatic hyperplasia. The section was incubated with ab318964 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K36) antibody [EPR28335-73] (AB318964)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling Histone H3 (di methyl K36) with ab318964 at 1/2000 (0.267 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human cerebrum. The section was incubated with ab318964 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IP

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Immunoprecipitation - Anti-Histone H3 (di methyl K36) antibody [EPR28335-73] (AB318964)

Histone H3 (di methyl K36) was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) histone lysate with ab318964 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab318964 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human cervical adenocarcinoma epithelial cell) histone lysate
Lane 2 : ab318964 IP in HeLa (human cervical adenocarcinoma epithelial cell) histone lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab318964 in HeLa histone lysate

All lanes:

Immunoprecipitation - Anti-Histone H3 (di methyl K36) antibody [EPR28335-73] (ab318964) at 1/30 dilution

All lanes:

HeLa (human cervical adenocarcinoma epithelial cell) histone lysate with NFDM/TBST

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 180s

• ChIP-seq

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ChIP-sequencing - Anti-Histone H3 (di methyl K36) antibody [EPR28335-73] (AB318964)

Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 8 µg of ab318964 [EPR28335-73]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 60 million reads. The Input control is also shown.

• ChIP-seq

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ChIP-sequencing - Anti-Histone H3 (di methyl K36) antibody [EPR28335-73] (AB318964)

Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 8 µg of ab318964 [EPR28335-73]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 60 million reads. The Input control is also shown.

• ChIP-seq

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ChIP-sequencing - Anti-Histone H3 (di methyl K36) antibody [EPR28335-73] (AB318964)

Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 8 µg of ab318964 [EPR28335-73]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 60 million reads. The Input control is also shown.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K36) antibody [EPR28335-73] (AB318964)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling Histone H3 (di methyl K36) with ab318964 at 1/2000 (0.267 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse cerebrum. The section was incubated with ab318964 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K36) antibody [EPR28335-73] (AB318964)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Histone H3 (di methyl K36) with ab318964 at 1/2000 (0.267 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen. The section was incubated with ab318964 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K36) antibody [EPR28335-73] (AB318964)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling Histone H3 (di methyl K36) with ab318964 at 1/2000 (0.267 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat spleen. The section was incubated with ab318964 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K36) antibody [EPR28335-73] (AB318964)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling Histone H3 (di methyl K36) with ab318964 at 1/2000 (0.267 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat cerebrum. The section was incubated with ab318964 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

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Western blot - Anti-Histone H3 (di methyl K36) antibody [EPR28335-73] (AB318964)

The identity of the bands higher than 16 kDa are unknown.

All lanes:

Western blot - Anti-Histone H3 (di methyl K36) antibody [EPR28335-73] (ab318964) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg with BSA/TBST

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg with BSA/TBST

Lane 3:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg with BSA/TBST

Lane 4:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg with BSA/TBST

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 16 kDa

false

Exposure time: 26s

• PepArr

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Peptide Array - Anti-Histone H3 (di methyl K36) antibody [EPR28335-73] (AB318964)

Peptide array analysis of ab318964 at 1 : 5000 (0.1 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Fluo 647nm conjugated at 1 : 50,000 dilution.