Anti-HDAC2 antibody [EPR20117] (ab219053)

Rabbit Recombinant Monoclonal HDAC2 antibody. Suitable for ChIC/CUT&RUN-seq, IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat samples. Cited in 1 publication.

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Images

Immunoprecipitation - Anti-HDAC2 antibody [EPR20117] (AB219053), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	ChIC/CUT&RUN-seq	IP	WB	ICC/IF	Flow Cyt (Intra)	IHC-P
Human	Tested	Tested	Tested	Tested	Expected	Tested
Mouse	Expected	Expected	Tested	Tested	Tested	Tested
Rat	Expected	Expected	Tested	Expected	Expected	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 5 µg	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/30	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/1000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes -
Species Human	Dilution info 1/1000	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/500	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Human, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/4000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/4000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/4000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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Target data

See full target information HDAC2

Function

Histone deacetylase that catalyzes the deacetylation of lysine residues on the N-terminal part of the core histones (H2A, H2B, H3 and H4) (PubMed:28497810). Histone deacetylation gives a tag for epigenetic repression and plays an important role in transcriptional regulation, cell cycle progression and developmental events (By similarity). Histone deacetylases act via the formation of large multiprotein complexes (By similarity). Forms transcriptional repressor complexes by associating with MAD, SIN3, YY1 and N-COR (PubMed:12724404). Component of a RCOR/GFI/KDM1A/HDAC complex that suppresses, via histone deacetylase (HDAC) recruitment, a number of genes implicated in multilineage blood cell development (By similarity). Acts as a component of the histone deacetylase NuRD complex which participates in the remodeling of chromatin (PubMed:16428440, PubMed:28977666). Component of the SIN3B complex that represses transcription and counteracts the histone acetyltransferase activity of EP300 through the recognition H3K27ac marks by PHF12 and the activity of the histone deacetylase HDAC2 (PubMed:37137925). Also deacetylates non-histone targets: deacetylates TSHZ3, thereby regulating its transcriptional repressor activity (PubMed:19343227). May be involved in the transcriptional repression of circadian target genes, such as PER1, mediated by CRY1 through histone deacetylation (By similarity). Involved in MTA1-mediated transcriptional corepression of TFF1 and CDKN1A (PubMed:21965678). In addition to protein deacetylase activity, also acts as a protein-lysine deacylase by recognizing other acyl groups: catalyzes removal of (2E)-butenoyl (crotonyl), lactoyl (lactyl) and 2-hydroxyisobutanoyl (2-hydroxyisobutyryl) acyl groups from lysine residues, leading to protein decrotonylation, delactylation and de-2-hydroxyisobutyrylation, respectively (PubMed:28497810, PubMed:29192674, PubMed:35044827).

Alternative names

Histone deacetylase 2, HD2, Protein deacylase HDAC2, HDAC2

Recommended products

Rabbit Recombinant Monoclonal HDAC2 antibody. Suitable for ChIC/CUT&RUN-seq, IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse, Rat samples. Cited in 1 publication.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR20117
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

HDAC2 or Histone Deacetylase 2 is an important enzyme that plays a role in chromatin remodeling and gene expression regulation. It removes acetyl groups from histone tails allowing chromatin to condense and silencing gene expression. HDAC2 has a molecular weight of approximately 55 kDa. It is expressed in various tissues including brain heart and skeletal muscles indicating its importance in different physiological areas. HDAC2 proteins are often studied using methods like HDAC2 ELISA and HDAC2 anticuero in research settings.

Biological function summary

HDAC2 is involved in cell cycle regulation and differentiation. It is part of the co-repressor complex and interacts with other proteins such as Sin3 and NuRD to repress transcription. These interactions are vital for maintaining normal cellular functions and ensuring proper response to external signals. This protein is also connected to HEK293T cells which are a common model in scientific studies due to their robust expression of proteins like HDAC2.

Pathways

HDAC2 participates in the regulation of transcriptional activity and cellular stress response. It is an important player in the histone modification pathway interacting with proteins such as HDAC1 and transcriptional regulators like REST. HDAC2 is also linked with the Notch signaling pathway which is essential for cell fate decisions. These pathways highlight its role in both maintaining cellular homeostasis and adapting to changes in the cellular environment.

Associated diseases and disorders

HDAC2 has a significant connection to neurodegenerative diseases and various cancers. Alzheimer's disease for instance shows altered expression levels of HDAC2 affecting synaptic plasticity and memory formation. In cancer HDAC2 interacts with oncogenic proteins such as p53 influencing tumor progression and metastasis. Monitoring HDAC2 levels and activity could provide insights into disease mechanisms and potential therapeutic targets.

Product promise

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In the unlikely event of one of our products not working as expected, you are covered by our product promise.

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21 product images

Immunoprecipitation - Anti-HDAC2 antibody [EPR20117] (ab219053)
HDAC2 was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab219053 at 1/30 dilution.
Western blot was performed from the immunoprecipitate using ab219053 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10000 dilution.
Lane 1: HeLa whole cell lysate 10 μg (Input).
Lane 2: ab219053 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab219053 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
All lanes: Immunoprecipitation - Anti-HDAC2 antibody [EPR20117] (ab219053)
Predicted band size: 55 kDa
Observed band size: 55 kDa
Western blot - Anti-HDAC2 antibody [EPR20117] (ab219053)
Lanes 1- 2: Merged signal (red and green). Green - ab219053 observed at 55 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.
ab219053 was shown to react with HDAC2 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line Human HDAC2 knockout HEK-293T cell line ab266589 (knockout cell lysate Human HDAC2 knockout HEK-293T cell lysate ab256938) was used. Wild-type HEK-293T and HDAC2 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab219053 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-HDAC2 antibody [EPR20117] (ab219053) at 1/1000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: HDAC2 knockout HEK-293T cell lysate at 20 µg
Lane 2: Western blot - Human HDAC2 knockout HEK-293T cell line (Human HDAC2 knockout HEK-293T cell line ab266589)
Performed under reducing conditions.
Predicted band size: 55 kDa
Observed band size: 55 kDa
Western blot - Anti-HDAC2 antibody [EPR20117] (ab219053)
Lanes 1-2: Merged signal (red and green). Green - ab219053 observed at 60 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
ab219053 Anti-HDAC2 antibody [EPR20117] was shown to specifically react with HDAC2 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line Human HDAC2 knockout HEK-293T cell line ab266588 (knockout cell lysate Human HDAC2 knockout HEK-293T cell lysate ab256937) was used. Wild-type and HDAC2 knockout samples were subjected to SDS-PAGE. ab219053 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 1000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-HDAC2 antibody [EPR20117] (ab219053) at 1/1000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: HDAC2 knockout HEK-293T cell lysate at 20 µg
Lane 2: Western blot - Human HDAC2 knockout HEK-293T cell line (Human HDAC2 knockout HEK-293T cell line ab266588)
Performed under reducing conditions.
Predicted band size: 55 kDa
Observed band size: 60 kDa
Western blot - Anti-HDAC2 antibody [EPR20117] (ab219053)
Lanes 1-2: Merged signal (red and green). Green - ab219053 observed at 60 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
ab219053 Anti-HDAC2 antibody [EPR20117] was shown to specifically react with HDAC2 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line Human HDAC2 knockout HEK-293T cell line ab266590 (knockout cell lysate Human HDAC2 knockout HEK-293T cell lysate ab256939) was used. Wild-type and HDAC2 knockout samples were subjected to SDS-PAGE. ab219053 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 1000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-HDAC2 antibody [EPR20117] (ab219053) at 1/1000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: HDAC2 knockout HEK-293T cell lysate at 20 µg
Lane 2: Western blot - Human HDAC2 knockout HEK-293T cell line (Human HDAC2 knockout HEK-293T cell line ab266590)
Performed under reducing conditions.
Predicted band size: 55 kDa
Observed band size: 60 kDa
Immunocytochemistry/ Immunofluorescence - Anti-HDAC2 antibody [EPR20117] (ab219053)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293 (Human epithelial cell line from embryonic kidney) cells labeling HDAC2 with ab219053 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing nuclear staining on HEK-293 cell line.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution.
Western blot - Anti-HDAC2 antibody [EPR20117] (ab219053)
Lanes 1 - 4: Merged signal (red and green). Green - ab219053 observed at 55 kDa. Red - loading control, Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484, observed at 37 kDa.
ab219053 was shown to specifically react with HDAC2 in wild type cells as signal was lost in HDAC2 knockout cells. Wild-type and HDAC2 knockout samples were subjected to SDS-PAGE. ab219053 and Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-HDAC2 antibody [EPR20117] (ab219053) at 1/1000 dilution
Lane 1: Wild-type HAP1 whole cell lysate at 20 µg
Lane 2: HDAC2 knockout HAP1 whole cell lysate at 20 µg
Lane 3: A431 whole cell lysate at 20 µg
Lane 4: HeLa whole cell lysate at 20 µg
Predicted band size: 55 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC2 antibody [EPR20117] (ab219053)
Immunohistochemical analysis of paraffin-embedded human testis tissue labeling HDAC2 with ab219053 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Nuclear staining on human testis is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-HDAC2 antibody [EPR20117] (ab219053)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-HDAC2 antibody [EPR20117] (ab219053) at 1/1000 dilution
Lane 1: His-tagged human HDAC2 recombinant protein (aa339-488) at 0.01 µg
Lane 2: His-tagged human HDAC1 recombinant protein (aa1-482) at 0.01 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 55 kDa
Observed band size: 22 kDa
Exposure time: 1s
Flow Cytometry (Intracellular) - Anti-HDAC2 antibody [EPR20117] (ab219053)
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling HDAC2 with ab219053 at 1/500 dilution (red) compared with a rabbit monoclonal IgG isotype control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.
Immunocytochemistry/ Immunofluorescence - Anti-HDAC2 antibody [EPR20117] (ab219053)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling HDAC2 with ab219053 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing nuclear staining on NIH/3T3 cell line.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC2 antibody [EPR20117] (ab219053)
Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling HDAC2 with ab219053 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Nuclear staining on mouse colon is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-HDAC2 antibody [EPR20117] (ab219053)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-HDAC2 antibody [EPR20117] (ab219053) at 1/5000 dilution
Lane 1: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2: SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate at 20 µg
Lane 3: HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 55 kDa
Observed band size: 55 kDa
Exposure time: 10s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC2 antibody [EPR20117] (ab219053)
Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling HDAC2 with ab219053 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Nuclear staining on rat spleen is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-HDAC2 antibody [EPR20117] (ab219053)
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1: 3 minutes; Lane 2: 15 seconds; Lane 3: 2 seconds.
All lanes: Western blot - Anti-HDAC2 antibody [EPR20117] (ab219053) at 1/5000 dilution
Lane 1: Human fetal brain lysate at 10 µg
Lane 2: Human fetal heart lysate at 10 µg
Lane 3: Human fetal kidney lysate at 10 µg
Secondary
All lanes: Western blot - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/4000 dilution
Predicted band size: 55 kDa
Observed band size: 55 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC2 antibody [EPR20117] (ab219053)
Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling HDAC2 with ab219053 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Nuclear staining on lymphocytes of human tonsil is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-HDAC2 antibody [EPR20117] (ab219053)
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1/2: 15 seconds; Lane 3: 30 seconds; Lane 4/5: 3 seconds; Lane 6/7: 1 second.
Lanes 1 - 5: Western blot - Anti-HDAC2 antibody [EPR20117] (ab219053) at 1/1000 dilution
Lanes 6 - 7: Western blot - Anti-HDAC2 antibody [EPR20117] (ab219053) at 1/5000 dilution
Lane 1: Mouse brain lysate at 10 µg
Lane 2: Mouse heart lysate at 10 µg
Lane 3: Rat heart lysate at 10 µg
Lane 4: Rat brain lysate at 10 µg
Lane 5: Rat spleen lysate at 10 µg
Lane 6: PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg
Lane 7: NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 10 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 11 kDa, 55 kDa
Observed band size: 55 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC2 antibody [EPR20117] (ab219053)
Immunohistochemical analysis of paraffin-embedded human prostate hyperplasia tissue labeling HDAC2 with ab219053 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Nuclear staining on luminal epithelial cells of human prostate hyperplasia; negative staining on basal cells.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC2 antibody [EPR20117] (ab219053)
Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue labeling HDAC2 with ab219053 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Nuclear Nuclear staining on tumor cells of prostate cancer; weak or negative staining on basal cells.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC2 antibody [EPR20117] (ab219053)
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling HDAC2 with ab219053 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Nuclear staining on tumor cells of human breast cancer is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC2 antibody [EPR20117] (ab219053)
Immunohistochemical analysis of paraffin-embedded human synovial sarcoma tissue labeling HDAC2 with ab219053 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Nuclear staining on human synovial sarcoma is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ChIC/CUT&RUN sequencing - Anti-HDAC2 antibody [EPR20117] (ab219053)
ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2 x 10^5 K-562 (Human chronic myelogenous leukemia lymphoblast) cells and 5µg of ab219053 [EPR20117]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 is also shown.

Additional screenshots of mapped reads can be found in the Protocol booklet in the Support and downloads section.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.