Anti-Dnmt3a (mutated R882H) antibody [EPR27389-11] - BSA and Azide free

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dnmt3a (mutated R882H) antibody [EPR27389-11] - BSA and Azide free (AB307541)

This data was developed using ab307540, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded HEK-293T cells tran tissue labeling Dnmt3a (mutated R882H) with ab307540 at 1/5000 (0.109 ug/ml) followed by a ready to use LeicaDS9800 (Bond Polymer Refine Detection) was used. Positive staining on HEK-293T cells transfected with a human DNMT3A (R882H) expression vector containing myc and his tag (Image A) and no staining on HEK-293 cells transfected with human wildtype DNMT3A expression vector containing myc and his tag (Image B).The section was incubated with ab307540 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) was used. Heat mediated antigen retrieval 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-Dnmt3a (mutated R882H) antibody [EPR27389-11] - BSA and Azide free (AB307541)

This data was developed using ab307540, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling Dnmt3a (mutated R882H) with ab307540 at 1/5000 (0.109 ug/ml) followed by a ready to use LeicaDS9800 (Bond Polymer Refine Detection) was used. Negative control : no staining on human testis.The section was incubated with ab307540 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) was used. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Dnmt3a (mutated R882H) antibody [EPR27389-11] - BSA and Azide free (AB307541)

This data was developed using ab307540, 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 labelling Dnmt3a (mutated R882H) with ab307540 at 1/1000 (0.546 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 showing positive staining in HEK-293T cells transfected with a human DNMT3A (R882H) expression vector containing a myc tag, and showing no staining in HEK-293 cells transfected with human wildtype DNMT3A 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 0.38 ug/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.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Dnmt3a (mutated R882H) antibody [EPR27389-11] - BSA and Azide free (AB307541)

This data was developed using ab307540, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HEK-293T (human embryonic kidney) cells transfected with a human DNMT3A expression vector containing a myc-his tag (Left) / HEK-293T transfected with a human DNMT3A (R882H) expression vector containing a myc-his tag (Right) cells labelling Dnmt3a (mutated R882H) with ab307540 at 1/500 dilution (0.1ug)/ Right and Left (Red) compared with a 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 1/2000 dilution was used as the secondary antibody.