Anti-Histone H4 (di methyl K20) antibody [EPR28338-29]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 (di methyl K20) antibody [EPR28338-29] (AB320813)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling Histone H4 with ab320813 at 1/100 (4.92 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human spleen. The section was incubated with ab320813 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

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 H4 (di methyl K20) antibody [EPR28338-29] (AB320813)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labeling Histone H4 with ab320813 at 1/100 (4.92 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 labeled 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 2.5ug/ml 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Histone H4 (di methyl K20) antibody [EPR28338-29] (AB320813)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling Histone H4 with ab320813 at 1/100 (4.92 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 NIH/3T3 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled 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 ug/ml 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 (di methyl K20) antibody [EPR28338-29] (AB320813)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling Histone H4 with ab320813 at 1/500 (0.984 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat spleen. The section was incubated with ab320813 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

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 H4 (di methyl K20) antibody [EPR28338-29] (AB320813)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Histone H4 with ab320813 at 1/500 (0.984 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen. The section was incubated with ab320813 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

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

Supplier Data

Western blot - Anti-Histone H4 (di methyl K20) antibody [EPR28338-29] (AB320813)

Blocking and diluting buffer and concentration : 1% BSA/TBST.

All lanes:

Western blot - Anti-Histone H4 (di methyl K20) antibody [EPR28338-29] (ab320813) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg

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: 11 kDa

false

Exposure time: 7s

• WB

Supplier Data

Western blot - Anti-Histone H4 (di methyl K20) antibody [EPR28338-29] (AB320813)

Blocking and diluting buffer and concentration : 1% BSA/TBST.

The identity of the bands higher than 15 kDa are unknown.

All lanes:

Western blot - Anti-Histone H4 (di methyl K20) antibody [EPR28338-29] (ab320813) at 1/1000 dilution

All lanes:

Human spleen tissue lysate at 20 µg

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: 11 kDa

false

Exposure time: 81s

• WB

Supplier Data

Western blot - Anti-Histone H4 (di methyl K20) antibody [EPR28338-29] (AB320813)

Blocking and diluting buffer and concentration : 1% BSA/TBST.

The identity of the bands higher than 15 kDa are unknown.

This blot was developed using a high sensitivity ECL substrate.

Exposure time : Lane 1-3 : 48 seconds; Lane 4-5 : 27 seconds

All lanes:

Western blot - Anti-Histone H4 (di methyl K20) antibody [EPR28338-29] (ab320813) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse testis tissue lysate at 20 µg

Lane 3:

Mouse spleen tissue lysate at 20 µg

Lane 4:

Rat brain tissue lysate at 20 µg

Lane 5:

Rat spleen tissue lysate at 20 µg

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: 11 kDa

true

• PepArr

Supplier Data

Peptide Array - Anti-Histone H4 (di methyl K20) antibody [EPR28338-29] (AB320813)

Peptide array analysis of ab320813 at 1 : 5000 (0.1 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Fluo 647nm conjugated at 1 : 50,000 dilution.

ab320813 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 the area under the curve when antibody binding values are plotted against the corresponding peptide concentration. Each circle area is normalized to the peptide with the strongest affinity.

Protein Array Washing Buffer, 2*10mins and 2*5mins

Please see the Product Protocol section for further details.

• Dot

Supplier Data

Dot Blot - Anti-Histone H4 (di methyl K20) antibody [EPR28338-29] (AB320813)

Dot blot analysis of Histone H4 using ab320813 at 1 : 1000 (0.492 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane1 : Histone H4 (di-methyl K20) peptide
Lane2 : Unmodified Histone H4 peptide
Lane3 : Histone H4 (mono-methyl K20) peptide
Lane4 : Histone H4 (tri-methyl K20) peptide

Exposure time : 3 minutes.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Dot Blot - Anti-Histone H4 (di methyl K20) antibody [EPR28338-29] (ab320813) at 1/1000 dilution

Lane 1:

Histone H4 (di-methyl K20) peptide

Lane 2:

Unmodified Histone H4 peptide

Lane 3:

Histone H4 (mono-methyl K20) peptide

Lane 4:

Histone H4 (tri-methyl K20) peptide

Secondary

All lanes:

Dot 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

false

Exposure time: 3min