All lanes : Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1000 dilution (unpurified)Lane 1 : HeLa cell lysates, untreated Lane 2 : HeLa cell lysates treated with TSALysates/proteins at 10 µg per lane.SecondaryHRP-labelled goat anti-rabbit at 1/2000 dilution
Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1000 dilution (unpurified) + HeLa cell lysate - treated with TSA at 10 µgSecondaryPeroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1500 dilution (purified) + HeLa cell lysate - treated with TSA at 10 µgSecondaryPeroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1000 dilution (unpurified) + C6 cell lysate - treated with TSA at 10 µgSecondaryPeroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1500 dilution (unpurified) + C6 cell lysate - treated with TSA at 10 µgSecondaryPeroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1000 dilution (purified) + Mouse spleen tissue lysate at 10 µgSecondaryPeroxidase-conjugated goat anti-rabbit IgG (H+L) at 1000
Anti-Histone H4 (acetyl K16) antibody [EPR1004] (ab109463) at 1/1500 dilution (purified) + Mouse spleen tissue lysate at 10 µgSecondaryPeroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue labelling Histone H4 (acetyl K16) with unpurified ab109463 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue labelling Histone H4 (acetyl K16) with purified ab109463 at 1/150. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human testis tissue labelling Histone H4 with unpurified ab109463 at 1/100.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human transitional cell carcinoma labelling Histone H4 (acetly K16) with unpurified ab109463 at 1/100.
Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Histone H4 (acetyl K16) with unpurified ab109463 (red) at 1/100. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.Control: primary antibody (1/100) and secondary antibody ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
Immunocytochemistry/Immunofluorescence analysis of HeLa cells treated with TSA labelling Histone H4 (acetyl K16) with unpurified ab109463 (red) at 1/100. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.Control: primary antibody (1/100) and secondary antibody ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Histone H4 (acetyl K16) with purified ab109463 (red) at 1/150. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.Control: primary antibody (1/150) and secondary antibody ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
Immunocytochemistry/Immunofluorescence analysis of HeLa cells treated with TSA labelling Histone H4 (acetyl K16) with purified ab109463 (red) at 1/150. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.Control: primary antibody (1/150) and secondary antibody ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
Overlay histogram showing HeLa cells stained with unpurified ab109463 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab109463, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Flow cytometry analysis of HeLa cells labelling Histone H4 (acetyl K16) with unpurified ab109463 (red) at 1/130. Cells were fixed with 80% methanol. A FITC-conjugated goat anti-rabbit IgG was used as the secondary antibody (1/150). A rabbit monoclonal IgG was used as the isotype control (green).
Flow cytometry analysis of HeLa cells labelling Histone H4 (acetyl K16) with purified ab109463 (red) at 1/200. Cells were fixed with 80% methanol. A FITC-conjugated goat anti-rabbit IgG was used as the secondary antibody (1/150). A rabbit monoclonal IgG was used as the isotype control (green).