Name: Anti-HDAC1 antibody [EPR23847-170]
Brand: RabMAb
SKU: ab280198
Rating: 4 (4 reviews)

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Western blot - Anti-HDAC1 antibody [EPR23847-170] (ab280198)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR23847-170] to HDAC1
Suitable for: Flow Cyt (Intra), ChIP, WB, IHC-P, ICC, IP
Knockout validated
Reacts with: Mouse, Rat, Human

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

Product name

Anti-HDAC1 antibody [EPR23847-170]
See all HDAC1 primary antibodies
Description

Rabbit monoclonal [EPR23847-170] to HDAC1
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt (Intra), ChIP, WB, IHC-P, ICC, IPmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human heart and kidney tissue lysates; Wild-type HAP1, Jurkat, K-562, HeLa, C6, RAW264.7, PC-12 and NIH/3T3 whole cell lysates; His-tagged human HDAC1 recombinant protein. IHC-P: Human tonsil and liver tissue; Mouse liver tissue; Rat liver tissue. ICC: HeLa and NIH/3T3 cells. Flow Cyt (intra): HeLa cells and NIH/3T3 cells. IP: HeLa and NIH/3T3 whole cell lysates. ChIP: Chromatin prepared from K-562 cells.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR23847-170
Isotype

IgG
Research areas

Alternative Versions
- Anti-HDAC1 antibody [EPR23847-170] - BSA and Azide free (ab280205)
Compatible Secondaries
Dyes/Markers
- Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)
HRP detection kit
- Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Related Products
- Anti-HDAC1 + HDAC2 antibody [EPR20327] (ab219054)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab280198 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
Flow Cyt (Intra)		1/500.
ChIP		Use 5 µg for 25 µg of chromatin.
WB	(4)	1/1000. Detects a band of approximately 62 kDa (predicted molecular weight: 55 kDa).
IHC-P		1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC		1/5000.
IP		1/30.

Notes
Flow Cyt (Intra) 1/500.
ChIP Use 5 µg for 25 µg of chromatin.
WB 1/1000. Detects a band of approximately 62 kDa (predicted molecular weight: 55 kDa).
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC 1/5000.
IP 1/30.

Function

Responsible for the deacetylation of lysine residues on the N-terminal part of the core histones (H2A, H2B, H3 and H4). Histone deacetylation gives a tag for epigenetic repression and plays an important role in transcriptional regulation, cell cycle progression and developmental events. Histone deacetylases act via the formation of large multiprotein complexes. Deacetylates SP proteins, SP1 and SP3, and regulates their function. Component of the BRG1-RB1-HDAC1 complex, which negatively regulates the CREST-mediated transcription in resting neurons. Upon calcium stimulation, HDAC1 is released from the complex and CREBBP is recruited, which facilitates transcriptional activation. Deacetylates TSHZ3 and regulates its transcriptional repressor activity. Deacetylates 'Lys-310' in RELA and thereby inhibits the transcriptional activity of NF-kappa-B.
Tissue specificity

Ubiquitous, with higher levels in heart, pancreas and testis, and lower levels in kidney and brain.
Sequence similarities

Belongs to the histone deacetylase family. HD type 1 subfamily.
Post-translational
modifications

Sumoylated on Lys-444 and Lys-476; which promotes enzymatic activity. Desumoylated by SENP1.
Phosphorylation on Ser-421 and Ser-423 promotes enzymatic activity and interactions with NuRD and SIN3 complexes.
Ubiquitinated by CHFR, leading to its degradation by the proteasome.
Cellular localization

Nucleus.
Target information above from: UniProt accession Q13547 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 3065 Human
- Entrez Gene: 433759 Mouse
- Entrez Gene: 297893 Rat
- Omim: 601241 Human
- SwissProt: Q13547 Human
- SwissProt: O09106 Mouse
- SwissProt: Q4QQW4 Rat
- Unigene: 88556 Human
- Unigene: 202504 Mouse
- Unigene: 391033 Mouse
- Unigene: 1863 Rat
see all
Alternative names
- DKFZp686H12203 antibody
- GON 10 antibody
- HD1 antibody
- HDAC 1 antibody
- HDAC1 antibody
- HDAC1_HUMAN antibody
- Histone deacetylase 1 antibody
- Reduced potassium dependency yeast homolog like 1 antibody
- RPD3 antibody
- RPD3L1 antibody
see all

Western blot - Anti-HDAC1 antibody [EPR23847-170] (ab280198)

All lanes : Anti-HDAC1 antibody [EPR23847-170] (ab280198) at 1/1000 dilution

Lane 1 : Wild-type HAP1 cell lysate at 20 µg
Lane 2 : HDAC1 knockout HAP1 cell lysate at 40 µg
Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 4 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (ab216773) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (ab216776) at 1/10000 dilution

Predicted band size: 55 kDa

ab280198 Anti-HDAC1 antibody [EPR23847-170] was shown to specifically react with HDAC1 in wild-type HAP1 cells. Loss of signal was observed when knockout cell line (knockout cell lysate) was used. Wild-type and HDAC1 knockout samples were subjected to SDS-PAGE. ab280198 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1/1000 dilution and 1/20000 dilution respectively. 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 1/20000 dilution for 1 hour at room temperature before imaging.
ChIP - Anti-HDAC1 antibody [EPR23847-170] (ab280198)

Chromatin was prepared from K-562 cells according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min.
The ChIP was performed with 25 µg of chromatin, 5 µg of ab280198 (red), or 5 µg of rabbit normal IgG ab172730 (gray) and 25 µl of Protein A/G Dynabeads. The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

*https://www.abcam.com/resources?keywords=X%20ChIP%20protocol
Immunoprecipitation - Anti-HDAC1 antibody [EPR23847-170] (ab280198)

HDAC1 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with ab280198 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab280198 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 ug

Lane 2: ab280198 IP in NIH/3T3 whole cell lysate 10 ug

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab280198 in NIH/3T3 whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 1 second.
Immunocytochemistry - Anti-HDAC1 antibody [EPR23847-170] (ab280198)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labelling HDAC1 with ab280198 at 1/5000 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear and weakly cytoplasmic 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 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 1/1000 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC1 antibody [EPR23847-170] (ab280198)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling HDAC1 with ab280198 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human tonsil (PMID:23109994). The section was incubated with ab280198 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Western blot - Anti-HDAC1 antibody [EPR23847-170] (ab280198)

All lanes : Anti-HDAC1 antibody [EPR23847-170] (ab280198) at 1/1000 dilution

Lane 1 : Human heart tissue lysate
Lane 2 : Human kidney tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution (VeriBlot for IP secondary antibody(HRP))

Predicted band size: 55 kDa
Observed band size: 62 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

The observed MW is consistent with what has been described in the literature (PMID:24551070).

Exposure time: 15 seconds.
Flow Cytometry (Intracellular) - Anti-HDAC1 antibody [EPR23847-170] (ab280198)

Flow cytometric analysis of 4% paraformaldehyde-fixe,d 90% methanol-permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling HDAC1 with ab280198 at 1/500 dilution (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.
Western blot - Anti-HDAC1 antibody [EPR23847-170] (ab280198)

All lanes : Anti-HDAC1 antibody [EPR23847-170] (ab280198) at 1/5000 dilution

Lane 1 : Jurkat (human T cell leukemia T lymphocyte) whole cell lysate
Lane 2 : K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate
Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 55 kDa
Observed band size: 62 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

The observed MW is consistent with what has been described in the literature (PMID: 24551070).

Exposure time: 37 seconds.
Immunocytochemistry - Anti-HDAC1 antibody [EPR23847-170] (ab280198)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 cells labelling HDAC1 with ab280198 at 1/5000 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear and weakly cytoplasmic 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 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 1/1000 dilution.
Western blot - Anti-HDAC1 antibody [EPR23847-170] (ab280198)

All lanes : Anti-HDAC1 antibody [EPR23847-170] (ab280198) at 1/1000 dilution

Lane 1 : His-tagged human HDAC1 recombinant protein (aa1-482)
Lane 2 : His-tagged human HDAC2 recombinant protein (aa1-488)

Lysates/proteins at 0.01 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 55 kDa
Observed band size: 66 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBST

This antibody has no cross-reaction with human HDAC2.

These recombinant proteins were made in house. These two recombinant proteins were expressed from E.coli expression systems.

Exposure time: 10 seconds.
Flow Cytometry (Intracellular) - Anti-HDAC1 antibody [EPR23847-170] (ab280198)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling HDAC1 with ab280198 at 1/500 dilution (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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC1 antibody [EPR23847-170] (ab280198)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling HDAC1 with ab280198 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human liver (PMID:18264140). The section was incubated with ab280198 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunoprecipitation - Anti-HDAC1 antibody [EPR23847-170] (ab280198)

HDAC1 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab280198 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab280198 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 ug

Lane 2: ab280198 IP in HeLa whole cell lysate 10 ug

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab280198 in HeLa whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 1 second.
Western blot - Anti-HDAC1 antibody [EPR23847-170] (ab280198)

All lanes : Anti-HDAC1 antibody [EPR23847-170] (ab280198) at 1/5000 dilution

Lane 1 : C6 (rat glial tumor glial cell) whole cell lysate
Lane 2 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 3 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
Lane 4 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 55 kDa
Observed band size: 62 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

The observed MW is consistent with what has been described in the literature (PMID:24551070).

Exposure time: 37 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC1 antibody [EPR23847-170] (ab280198)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling HDAC1 with ab280198 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on mouse liver. The section was incubated with ab280198 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC1 antibody [EPR23847-170] (ab280198)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling HDAC1 with ab280198 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on rat liver. The section was incubated with ab280198 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheet download

Download

Publishing research using ab280198? Please let us know so that we can cite the reference in this datasheet.

ab280198 has not yet been referenced specifically in any publications.

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Application

Western blot

Sample

Human Cell lysate - whole cell (Lung cell line)

Gel Running Conditions

Reduced Denaturing (10)

Loading amount

89 µg

Specification

Lung cell line

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Dec 14 2021

Application

Western blot

Sample

Mouse Cell lysate - whole cell (Leukemia T cell line)

Gel Running Conditions

Reduced Denaturing (10)

Loading amount

89 µg

Specification

Leukemia T cell line

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Dec 14 2021

Application

Western blot

Sample

Human Cell lysate - whole cell (cell line)

Gel Running Conditions

Reduced Denaturing (10%)

Loading amount

45 µg

Specification

cell line

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Dec 14 2021

Application

Western blot

Sample

Mouse Cell lysate - whole cell (Cell line)

Gel Running Conditions

Reduced Denaturing (10%)

Loading amount

45 µg

Specification

Cell line

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Dec 14 2021

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Dyes/Markers

HRP detection kit

Isotype control

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

Certificate of Compliance

References (0)

Customer reviews and Q&As

Western blot abreview for Anti-HDAC1 antibody [EPR23847-170]

Western blot abreview for Anti-HDAC1 antibody [EPR23847-170]

Western blot abreview for Anti-HDAC1 antibody [EPR23847-170]

Western blot abreview for Anti-HDAC1 antibody [EPR23847-170]

Post-translational
modifications