Alexa Fluor® 647 Anti-Histone H3 antibody [EPR16987] - Nuclear Marker

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-Histone H3 antibody [EPR16987] - Nuclear Marker (AB207543)

Overlay histogram showing HeLa cells stained with ab207543 (red line). The cells were fixed with 80 % methanol (5 min) and then permeabilized with 0.1 % PBS-Triton X-100 for 15 min. The cells were incubated in 1x PBS containing 10 % normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab207543) (1x 10⁶ in 100 μL at 1 μg/ml (1/500)) for 30 min at 22°C. Isotype control antibody (black line) was Rabbit IgG (monoclonal) Alexa Fluor^®647 (ab199093) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 40 mW Red laser (640nm) and 670/14 bandpass filter.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Histone H3 antibody [EPR16987] - Nuclear Marker (AB207543)

ab207543 staining Histone H3 - Nuclear Loading Control and ChIP Grade in HeLa cells. The cells were fixed with 100% methanol (5 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 ab207543 at 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/250 dilution (shown in green). Nuclear DNA was labeled with DAPI (shown in blue).

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 647 Anti-Histone H3 antibody [EPR16987] - Nuclear Marker (AB207543)

Immunohistochemistry analysis of paraffin-embedded human colon tissue sections labeling Histone H3 with ab207543 at 1/100, incubated with ab207543 for 60 mins at room temperature (shown in red). Nuclear DNA was labeled with DAPI (shown in blue). The section was then mounted using Fluoromount®. Antigen retrieval was heat mediated with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 40 mins. Nuclear staining on human colon. The immunostaining was performed on a Leica Biosystems BOND RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-Histone H3 antibody [EPR16987] - Nuclear Marker (AB207543)

Overlay histogram showing NIH/3T3 (mouse embryonic fibroblast) cells stained with ab207543 at 1/500 (red line). The cells were fixed with 4% paraformaldehyde and then permeabilized with 90% methanol. Isotype control antibody (black line) was Rabbit IgG (monoclonal) Alexa Fluor®647 (ab199093). Unlabelled sample, cells without incubation with primary antibody and secondary antibody (blue line) was also used as a control.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 647 Anti-Histone H3 antibody [EPR16987] - Nuclear Marker (AB207543)

Immunohistochemistry analysis of paraffin-embedded mouse liver tissue sections labeling Histone H3 with ab207543 at 1/100, incubated with ab207543 for 60 mins at room temperature (shown in red). Nuclear DNA was labeled with DAPI (shown in blue). The section was then mounted using Fluoromount®. Antigen retrieval was heat mediated with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 40 mins. Nuclear staining on mouse liver. The immunostaining was performed on a Leica Biosystems BOND RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).