Anti-HDAC1 antibody [EPR460(2)] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC1 antibody [EPR460(2)] - BSA and Azide free (AB213701)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109411).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-HDAC1 antibody [EPR460(2)] - BSA and Azide free (AB213701)

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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109411).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-HDAC1 antibody [EPR460(2)] - BSA and Azide free (AB213701)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109411).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-HDAC1 antibody [EPR460(2)] - BSA and Azide free (AB213701)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109411).

Immunocytochemical analysis of 4% paradormaldehyde fixed, 0.1% Triton X-100 permeabilised HeLa cells labeling HDAC1 with ab109411 at 1/50 (10.92 μg/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) secondary at 1/1000 (2 μg/ml) dilution. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain at 1/200 (2.5 μg/ml) dilution. Nuclear counterstain : DAPI.

Confocal image showing mainly nuclear staining in HeLa cell line

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC1 antibody [EPR460(2)] - BSA and Azide free (AB213701)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109411).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-HDAC1 antibody [EPR460(2)] - BSA and Azide free (AB213701)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109411).

Immunocytochemical analysis of 4% paradormaldehyde fixed, 0.1% Triton X-100 permeabilised NIH/3T3 cells labeling HDAC1 with ab109411 at 1/50 (10.92 μg/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) secondary at 1/1000 (2 μg/ml) dilution. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain at 1/200 (2.5 μg/ml) dilution. Nuclear counterstain : DAPI.

Confocal image showing nuclear staining in NIH/3T3 cell line

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC1 antibody [EPR460(2)] - BSA and Azide free (AB213701)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109411).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-HDAC1 antibody [EPR460(2)] - BSA and Azide free (AB213701)

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.<\p>

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109411).

• WB

Supplier Data

Western blot - Anti-HDAC1 antibody [EPR460(2)] - BSA and Azide free (AB213701)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109411).

All lanes:

Western blot - Anti-HDAC1 antibody [EPR460(2)] - BSA and Azide free (ab213701)

Predicted band size: 55 kDa

false

• WB

Supplier Data

Western blot - Anti-HDAC1 antibody [EPR460(2)] - BSA and Azide free (AB213701)

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.

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109411).

All lanes:

Western blot - Anti-HDAC1 antibody [EPR460(2)] - BSA and Azide free (ab213701)

Predicted band size: 55 kDa

false

• WB

Unknown

Western blot - Anti-HDAC1 antibody [EPR460(2)] - BSA and Azide free (AB213701)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109411).

All lanes:

Western blot - Anti-HDAC1 antibody [EPR460(2)] - Nuclear Loading Control (<a href='/en-us/products/primary-antibodies/hdac1-antibody-epr4602-nuclear-loading-control-ab109411'>ab109411</a>) 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

• WB

CiteAb

Western blot - Anti-HDAC1 antibody [EPR460(2)] - BSA and Azide free (AB213701)

HDAC1 western blot using anti-HDAC1 antibody [EPR460(2)] - BSA and Azide free ab213701. Publication image and figure legend from Lin, M. Y., de Zoete, M. R., et al., 2015, Front Immunol, PubMed 26579129.

ab213701 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab213701 please see the product overview.

Immunomodulatory effects of SCFAs are mimicked by the histone deacetylase inhibitor TSA. (A) HDAC activity in HeLa lysates in the presence of 10 mM Na-butyrate, Na-propionate or 2 μM TSA. (B) Immunoblots of HEK293 and HeLa 57A incubated with 10 mM Na-Bu, 10 mM Na-Pro or 2 μM TSA for 30 min or 5 h analyzed with anti-acetyl-lysine antibody to visualize total histone acetylation and anti-HDAC1, HDAC2, HDAC3, and GAPDH antibodies. (C,D). Immunomodulatory effects of SCFAs are mimics by histone deacetylase inhibitor TSA. TLR5-NF-κB assays of HEK293 or HeLa 57A expressing TLR5 and an NF-κB-luciferase reporter. Cells were incubated with 10 mM Na-Bu, 10 mM Na-Pro or 2 μM TSA for 30 min followed by stimulation with S. enteriditis flagellin for 5 h. (E) TLR4-NF-κB assays of HEK293 or HeLa 57A expressing TLR4 and an NF-κB-luciferase reporter incubated with SCFA and LPS. Bars depict mean and SEM of three independent experiments. p < 0.05 (*), p < 0.01 (**), p < 0.005 (***).

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