Recombinant Anti-HDAC1 antibody [EPR460(2)] (ab109411)

Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 2ug/ml dilution.

Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 2ug/ml dilution.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse hippocampus tissue labeling HDAC1 with ab109411 at 1/500 (5.55 ug/ml) dilution followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 ug/ml) dilution (Green). Nuclear staining on mouse hippocampus. is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1000 (2 ug/ml) dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labelling HDAC1 with Purified ab109411 at 1:20 dilution (5 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150081) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

Immunocytochemistry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling HDAC1 with Purified ab109411 at 1:50 dilution (2.4 µg/ml). Cells were fixed in 100% Methanol and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat spleen tissue sections labeling HDAC1 with Purified ab109411 at 1:100 dilution (1.2 µg/ml). Heat mediated antigen retrieval was performed using . Tissue was counterstained with Hematoxylin. Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control. 

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse spleen tissue sections labeling HDAC1 with Purified ab109411 at 1:100 dilution (1.2 µg/ml). Heat mediated antigen retrieval was performed using . Tissue was counterstained with Hematoxylin. Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control. 

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue sections labeling HDAC1 with Purified ab109411 at 1:100 dilution (1.2 µg/ml). Heat mediated antigen retrieval was performed using . Tissue was counterstained with Hematoxylin. Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control. 

Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: HDAC1 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: Human breast carcinoma lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab109411 observed at 65 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab109411 was shown to recognize HDAC1 when HDAC1 knockout samples were used, along with additional cross-reactive bands. Wild-type and HDAC1 knockout samples were subjected to SDS-PAGE. ab109411 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed withGoat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 h at room temperature before imaging

Immunohistochemical analysis of 4% PFA fixed, 0.2% Triton X-100 permeabilised Mouse hippocampus staining HDAC1 with ab109411 at 1/500 dilution (5.55 μg/ml). ab150077 AlexaFluor®488 Goat anti-Rabbit was used as a secondary at 1/1000 (2 μg/ml) dilution. Nuclear counterstain: DAPI. 

Nuclear staining on mouse hippocampus.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)

Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: HDAC1 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: Human breast carcinoma lysate (20 µg) or K562 lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab109411 observed at 65 kDa. Red - loading control, ab8245, observed at 37 kDa.

This western blot image is a comparison between ab109411 and a competitor's top cited rabbit polyclonal antibody.