Recombinant Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR5001] to HDAC2 - ChIP Grade
- Suitable for: WB, ICC/IF, ChIP
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-HDAC2 antibody [EPR5001] - ChIP Grade
See all HDAC2 primary antibodies -
Description
Rabbit monoclonal [EPR5001] to HDAC2 - ChIP Grade -
Host species
Rabbit -
Tested applications
Suitable for: WB, ICC/IF, ChIPmore details
Unsuitable for: Flow Cyt or IHC-P -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Rat -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Wild-type HEK-293T, HAP1, HeLa, A431, SH-SY5Y, and Jurkat cell lysates. ICC/IF: Wild-type HAP1 cells. ChIP: Chromatin extract from HeLa cells.
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.2
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR5001 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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ChIP Related Products
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab124974 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 |
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WB | (3) |
1/10000 - 1/50000. Predicted molecular weight: 55 kDa.
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ICC/IF |
Use a concentration of 0.5 µg/ml.
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ChIP | (1) |
Use 10 µg for 25 µg of chromatin.
Please note product formulation is not optimised for ChIP application |
Notes |
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WB
1/10000 - 1/50000. Predicted molecular weight: 55 kDa. |
ICC/IF
Use a concentration of 0.5 µg/ml. |
ChIP
Use 10 µg for 25 µg of chromatin. Please note product formulation is not optimised for ChIP application |
Target
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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.
Forms transcriptional repressor complexes by associating with MAD, SIN3, YY1 and N-COR. Interacts in the late S-phase of DNA-replication with DNMT1 in the other transcriptional repressor complex composed of DNMT1, DMAP1, PCNA, CAF1. Deacetylates TSHZ3 and regulates its transcriptional repressor activity. -
Tissue specificity
Widely expressed; lower levels in brain and lung. -
Sequence similarities
Belongs to the histone deacetylase family. HD type 1 subfamily. -
Post-translational
modificationsS-nitrosylated by GAPDH. In neurons, S-Nitrosylation at Cys-262 and Cys-274 does not affect the enzyme activity but abolishes chromatin-binding, leading to increases acetylation of histones and activate genes that are associated with neuronal development. In embryonic cortical neurons, S-Nitrosylation regulates dendritic growth and branching. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 3066 Human
- Entrez Gene: 15182 Mouse
- Entrez Gene: 84577 Rat
- Omim: 605164 Human
- SwissProt: Q92769 Human
- SwissProt: P70288 Mouse
- Unigene: 3352 Human
- Unigene: 19806 Mouse
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Alternative names
- D10Wsu179e antibody
- HD 2 antibody
- HD2 antibody
see all
Images
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All lanes : Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974) at 1/10000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : HDAC2 knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 55 kDa
Observed band size: 55 kDaLanes 1- 2: Merged signal (red and green). Green - ab124974 observed at 55 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab124974 was shown to react with HDAC2 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266589 (knockout 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. ab124974 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 10000 dilution and a 1 in 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 in 20000 dilution for 1 hour at room temperature before imaging.
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ab124974 staining HDAC2 in wild-type HAP1 cells (top panel) and HDAC2 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab124974 at 0.5μg/ml and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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All lanes : Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974) at 1/10000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : HDAC2 knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 55 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?Lanes 1-2: Merged signal (red and green). Green - ab124974 observed at 60 kDa. Red - loading control ab8245 observed at 37 kDa.
ab124974 Anti-HDAC2 antibody [EPR5001] was shown to specifically react with HDAC2 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266590 (knockout cell lysate ab256939) was used. Wild-type and HDAC2 knockout samples were subjected to SDS-PAGE. ab124974 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 10000 and 1 in 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 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974) at 1/10000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : HDAC2 knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 55 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?Lanes 1-2: Merged signal (red and green). Green - ab124974 observed at 60 kDa. Red - loading control ab8245 observed at 37 kDa.
ab124974 Anti-HDAC2 antibody [EPR5001] was shown to specifically react with HDAC2 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266588 (knockout cell lysate ab256937) was used. Wild-type and HDAC2 knockout samples were subjected to SDS-PAGE. ab124974 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 10000 Dilution and 1 in 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 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974) at 1/10000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : HDAC2 knockout HAP1 whole cell lysate
Lane 3 : A431 whole cell lysate
Lane 4 : HeLa whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 55 kDaLanes 1 - 4: Merged signal (red and green). Green - ab124974 observed at 60 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab124974 was shown to recognize HDAC2 when HDAC2 knockout samples were used, along with additional cross-reactive bands. Wild-type and HDAC2 knockout samples were subjected to SDS-PAGE. ab124974 and ab8245 (Mouse anti GAPDH loading control) were but diluted at 1/10000 and incubated overnight at 4°C. Blots were developed with 800CW Goat anti Rabbit and 680CW Goat anti Mouse secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974) at 1/10000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : SH-SY5Y cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : A431 cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 55 kDa -
Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 10µg of ab124974 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach). Primers and probes are located in the first kb of the transcribed region.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (10)
ab124974 has been referenced in 10 publications.
- Liu Z et al. nMOWChIP-seq: low-input genome-wide mapping of non-histone targets. NAR Genom Bioinform 4:lqac030 (2022). PubMed: 35402909
- Chen Z et al. Imaging assisted evaluation of antitumor efficacy of a new histone deacetylase inhibitor in the castration-resistant prostate cancer. Eur J Nucl Med Mol Imaging 48:53-66 (2021). PubMed: 32592040
- Xie L et al. Elucidation of the Hdac2/Sp1/miR-204-5p/Bcl-2 axis as a modulator of cochlear apoptosis via in vivo/in vitro models of acute hearing loss. Mol Ther Nucleic Acids 23:1093-1109 (2021). PubMed: 33614251
- Pan L et al. Silencing of CREB Inhibits HDAC2/TLR4/NF-?B Cascade to Relieve Severe Acute Pancreatitis-Induced Myocardial Injury. Inflammation 44:1565-1580 (2021). PubMed: 33725236
- Xu L et al. H3K14 hyperacetylation-mediated c-Myc binding to the miR-30a-5p gene promoter under hypoxia postconditioning protects senescent cardiomyocytes from hypoxia/reoxygenation injury. Mol Med Rep 23:N/A (2021). PubMed: 33880587
- Gilbert TM et al. Neuroepigenetic signatures of age and sex in the living human brain. Nat Commun 10:2945 (2019). PubMed: 31270332
- Link S et al. PWWP2A binds distinct chromatin moieties and interacts with an MTA1-specific core NuRD complex. Nat Commun 9:4300 (2018). PubMed: 30327463
- Hanigan TW et al. Scaffold dependent histone deacetylase (HDAC) inhibitor induced re-equilibration of the subcellular localization and post-translational modification state of class I HDACs. PLoS One 12:e0186620 (2017). PubMed: 29045501
- Hanigan TW et al. Divergent JNK Phosphorylation of HDAC3 in Triple-Negative Breast Cancer Cells Determines HDAC Inhibitor Binding and Selectivity. Cell Chem Biol 24:1356-1367.e8 (2017). PubMed: 28943357
- Lin MY et al. Redirection of Epithelial Immune Responses by Short-Chain Fatty Acids through Inhibition of Histone Deacetylases. Front Immunol 6:554 (2015). WB ; Human . PubMed: 26579129