Anti-HDAC1 antibody [EPR460(2)] - Nuclear Loading Control

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-HDAC1 antibody [EPR460(2)] - Nuclear Loading Control (AB109411)

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).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-HDAC1 antibody [EPR460(2)] - Nuclear Loading Control (AB109411)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling HDAC1 with ab109411 at 1/50 (10.92 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing mainly nuclear staining in HeLa cell line ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 2ug/ml dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC1 antibody [EPR460(2)] - Nuclear Loading Control (AB109411)

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.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-HDAC1 antibody [EPR460(2)] - Nuclear Loading Control (AB109411)

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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC1 antibody [EPR460(2)] - Nuclear Loading Control (AB109411)

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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC1 antibody [EPR460(2)] - Nuclear Loading Control (AB109411)

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.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-HDAC1 antibody [EPR460(2)] - Nuclear Loading Control (AB109411)

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).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-HDAC1 antibody [EPR460(2)] - Nuclear Loading Control (AB109411)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 cells labelling HDAC1 with ab109411 at 1/50 (10.92 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing nuclear staining in NIH/3T3 cell line ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 2ug/ml dilution.

• WB

Lab

Western blot - Anti-HDAC1 antibody [EPR460(2)] - Nuclear Loading Control (AB109411)

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.

All lanes:

Western blot - Anti-HDAC1 antibody [EPR460(2)] - Nuclear Loading Control (ab109411)

Predicted band size: 41 kDa,55 kDa,88 kDa

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• WB

Lab

Western blot - Anti-HDAC1 antibody [EPR460(2)] - Nuclear Loading Control (AB109411)

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

All lanes:

Western blot - Anti-HDAC1 antibody [EPR460(2)] - Nuclear Loading Control (ab109411)

Predicted band size: 55 kDa

false

• WB

Unknown

Western blot - Anti-HDAC1 antibody [EPR460(2)] - Nuclear Loading Control (AB109411)

All lanes:

Western blot - Anti-HDAC1 antibody [EPR460(2)] - Nuclear Loading Control (ab109411) at 1/1000 dilution

Lane 1:

Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate at 20 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 4:

C6 (Rat glial tumor glial cell) 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 1/20000 dilution

Predicted band size: 55 kDa

false