Anti-HDAC1 antibody [EPR5517(2)] - Nuclear Loading Control

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC1 antibody [EPR5517(2)] - Nuclear Loading Control (AB150399)

Immunohistochemical analysis of paraffin embedded normal Human tonsil tissue using ab150399 showing +ve staining.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC1 antibody [EPR5517(2)] - Nuclear Loading Control (AB150399)

Immunohistochemical analysis of paraffin embedded normal Human stomach tissue using ab150399 showing +ve staining.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC1 antibody [EPR5517(2)] - Nuclear Loading Control (AB150399)

Immunohistochemical analysis of paraffin embedded Human Ovarian carcinoma tissue using ab150399 showing +ve staining.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC1 antibody [EPR5517(2)] - Nuclear Loading Control (AB150399)

Immunohistochemical analysis of paraffin embedded Human Thyroid gland carcinoma tissue using ab150399 showing +ve staining.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC1 antibody [EPR5517(2)] - Nuclear Loading Control (AB150399)

Immunohistochemical analysis of paraffin embedded normal Human colon tissue using ab150399 showing +ve staining.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-HDAC1 antibody [EPR5517(2)] - Nuclear Loading Control (AB150399)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling HDAC1 with ab150399 at 1/100 (5.17 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 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.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-HDAC1 antibody [EPR5517(2)] - Nuclear Loading Control (AB150399)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling HDAC1 with ab150399 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC1 antibody [EPR5517(2)] - Nuclear Loading Control (AB150399)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labelling HDAC1 with ab150399 at 1/50 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IP

Lab

Immunoprecipitation - Anti-HDAC1 antibody [EPR5517(2)] - Nuclear Loading Control (AB150399)

Purified ab150399 at 1/30 dilution (2μg) immunoprecipitating HDAC1 in Jurkat whole cell lysate.
Lane 1 (input) : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10μg
Lane 2 (+) : ab150399 + Jurkat whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab150399 in Jurkat whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 62 kDa
Faint band above 62kDa could be Sumoylated HDAC1. (PMID : 28186506)

All lanes:

Immunoprecipitation - Anti-HDAC1 antibody [EPR5517(2)] - Nuclear Loading Control (ab150399)

Predicted band size: 55 kDa

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• WB

Lab

Western blot - Anti-HDAC1 antibody [EPR5517(2)] - Nuclear Loading Control (AB150399)

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 - ab150399 observed at 65 kDa. Red - loading control, ab8245, observed at 37 kDa.

This western blot image is a comparison between ab150399 and a competitor's top cited rabbit polyclonal antibody.

All lanes:

Western blot - Anti-HDAC1 antibody [EPR5517(2)] - Nuclear Loading Control (ab150399)

Predicted band size: 55 kDa

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• WB

Lab

Western blot - Anti-HDAC1 antibody [EPR5517(2)] - Nuclear Loading Control (AB150399)

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 - ab150399 observed at 65 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab150399 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. ab150399 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776)
secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-HDAC1 antibody [EPR5517(2)] - Nuclear Loading Control (ab150399)

Predicted band size: 55 kDa

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• WB

Unknown

Western blot - Anti-HDAC1 antibody [EPR5517(2)] - Nuclear Loading Control (AB150399)

All lanes:

Western blot - Anti-HDAC1 antibody [EPR5517(2)] - Nuclear Loading Control (ab150399) at 1/1000 dilution

Lane 1:

K562 cell lysate at 10 µg

Lane 2:

Jurkat cell lysate at 10 µg

Lane 3:

MCF7 cell lysate at 10 µg

Lane 4:

HeLa cell lysate at 10 µg

Predicted band size: 55 kDa

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• ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Anti-HDAC1 antibody [EPR5517(2)] - Nuclear Loading Control (AB150399)

CUT&RUN was performed using the ChIC/CUT&RUN pAG-MNAse ab285373 2.5 x 10^5 HeLa cells and 5μg of ab150399 [EPR5517(2)]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. Additional screenshots of mapped reads can be downloaded here.

• ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Anti-HDAC1 antibody [EPR5517(2)] - Nuclear Loading Control (AB150399)

ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2 x 10^5 HeLa (Human cervix adenocarcinoma epithelial cell line) cells and 5 µg of ab150399 [EPR5517(2)]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. Additional screenshots of mapped reads can be found in the Protocol booklet in the Product Protocol section. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• WB

CiteAb

Western blot - Anti-HDAC1 antibody [EPR5517(2)] - Nuclear Loading Control (AB150399)

HDAC1 western blot using anti-HDAC1 antibody [EPR5517(2)] ab150399. Publication image and figure legend from Dong, L. H., Cheng, S., et al., 2013, J Hematol Oncol, PubMed 23866964.

ab150399 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 ab150399 please see the product overview.

Valproic acid potentiated temsirolimus to trigger Burkitt leukemia/lymphoma (BL) cell autophagy through inhibiting HDAC1. (A) Valproic acid (VPA), either alone or in combination, inhibited HDAC1 expression (upper panel), in parallel with decreased HDAC1 enzymatic activity (lower panel), but increased CDKN1A and CDKN1B expression (upper panel). (B) Comparing with the negative control (CON siRNA), VPA failed to alter CDKN1A, CDKN1B, or LC3-I/II expression in Namalwa cells transfected with HDAC1 siRNA. (C and D) HDAC1 siRNA abrogated VPA-induced inhibition of tumor cell growth (C, *P<0.05 comparing with CON siRNA) and induction of autophagy (D, as detected by immunefluorescence study on BECN1 and P62).

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