Anti-Histone H2B (mutated E76Q) antibody [EPR27299-85] - BSA and Azide free

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Histone H2B (mutated E76Q) antibody [EPR27299-85] - BSA and Azide free (AB309101)

This data was developed using ab309100, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype control (Left) / 293T cells transfected with a human H2BC4(E76Q) expression vector containing a myc tag (Middle) / 293T cells transfected with a human H2BC4 (WT) expression vector containing a myc tag (Right) cells labeling Histone H2B (mutated E76Q) with ab309100 at 1/500 dilution (0.1 ug)/Right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Histone H2B (mutated E76Q) antibody [EPR27299-85] - BSA and Azide free (AB309101)

This data was developed using ab309100, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labeling Histone H2B (mutated E76Q) with ab309100 at 1/1000 (0.513 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 positive staining in 293T cells transfected with a human H2BC4 E76Q expression vector containing a myc tag, while showing no staining in 293T cells transfected with a human H2BC4 (WT) expression vector containing a myc tag. The image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). is observed. Myc-Tag Mouse mab (Alexa Fluor® 647) was used to counterstain tubulin at 1/100 0.38ug/ml dilution (Red). 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2B (mutated E76Q) antibody [EPR27299-85] - BSA and Azide free (AB309101)

This data was developed using ab309100, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling Histone H2B (mutated E76Q) with ab309100 at 1/5000 (0.103 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining on human cerebrum. The section was incubated with ab309100 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2B (mutated E76Q) antibody [EPR27299-85] - BSA and Azide free (AB309101)

This data was developed using ab309100, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human tissue labeling Histone H2B (mutated E76Q) with ab309100 at 1/5000 (0.103 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on (B) HEK-293T transfected with a human H2BC4 E76Q expression vector containing a His-tag cell pellet. No staining on (A) HEK-293T transfected with a human H2BC4 (WT) expression vector containing a His-tag cell pellet and (C) HEK-293T transfected with an empty vector containing a His-tag cell pellet. The section was incubated with ab309100 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2B (mutated E76Q) antibody [EPR27299-85] - BSA and Azide free (AB309101)

This data was developed using ab309100, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling Histone H2B (mutated E76Q) with ab309100 at 1/5000 (0.103 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining on rat cerebrum. The section was incubated with ab309100 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2B (mutated E76Q) antibody [EPR27299-85] - BSA and Azide free (AB309101)

This data was developed using ab309100, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling Histone H2B (mutated E76Q) with ab309100 at 1/5000 (0.103 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining on mouse cerebrum. The section was incubated with ab309100 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

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Western blot - Anti-Histone H2B (mutated E76Q) antibody [EPR27299-85] - BSA and Azide free (AB309101)

This data was developed using ab309100, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST In Western blot, anti-His antibody (ab213204) staining at 1/5000 dilution. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Histone H2B (mutated E76Q) antibody [EPR27299-85] (<a href='/en-us/products/primary-antibodies/histone-h2b-mutated-e76q-antibody-epr27299-85-ab309100'>ab309100</a>) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containi a His-tag whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human H2BC4 E76Q expression vector containi a His-tag whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human H2BC4 (WT) expression vector containi a His-tag 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: 20 kDa

false

Exposure time: 26s

• Dot

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Dot Blot - Anti-Histone H2B (mutated E76Q) antibody [EPR27299-85] - BSA and Azide free (AB309101)

This data was developed using ab309100, the same antibody clone in a different buffer formulation. Dot blot analysis of Histone H2B (mutated E76Q) using ab309100 at 1 : 1000 (0.513 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution. Exposure time : 180 seconds Blocking and diluting buffer and concentration : 5% NFDM/TBST