Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y461] to HDAC2
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
参阅全部 HDAC2 一抗
描述兔单克隆抗体[Y461] to HDAC2
特异性ab32117 recognises HDAC2.
经测试应用适用于: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details
种属反应性与反应: Mouse, Rat, Human
Synthetic peptide within Human HDAC2 aa 450-550 (C terminal). The exact sequence is proprietary.
- WB: HAP1, A431, Hela and K562 cell lysate and rat brain tissue homogenate. IHC-P: Human breast carcinoma and rat spinal cord tissue. ICC/IF: MCF-7 and wildtype HAP1 cells. Flow Cyt: HeLa cells.
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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
存放说明Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
- Anti-HDAC2 antibody [Y461] (HRP) (ab195851)
- Anti-HDAC2 antibody [Y461] (Alexa Fluor® 488) (ab196471)
- Anti-HDAC2 antibody [Y461] (Alexa Fluor® 647) (ab196518)
- Anti-HDAC2 antibody [Y461] (PE) (ab210827)
- Anti-HDAC2 antibody [Y461] - BSA and Azide free (ab213700)
- Anti-HDAC2 antibody [Y461] (Alexa Fluor® 568) (ab214384)
Our Abpromise guarantee covers the use of ab32117 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/2000. Predicted molecular weight: 55 kDa.|
|IHC-P||Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|ICC/IF||1/250 - 1/500.|
|Flow Cyt||1/60 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
功能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.
组织特异性Widely expressed; lower levels in brain and lung.
序列相似性Belongs to the histone deacetylase family. HD type 1 subfamily.
翻译后修饰S-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.
- Information by UniProt
- D10Wsu179e antibody
- HD 2 antibody
- HD2 antibody
Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: HDAC2 knockout HAP1 whole cell lysate (20 µg)
Lane 3: A431 whole cell lysate (20 µg)
Lane 4: Hela whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab32117 observed at 60 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab32117 was shown to specifically react with HDAC2 when HDAC2 knockout samples were used. Wild-type and HDAC2 knockout samples were subjected to SDS-PAGE. ab32117 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/2000 and 1/10000 respectively. 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.
ab32117 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 ab32117 at 1/250 dilution 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).
ab32117 staining HDAC2 in MCF-7 (human breast carcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. DAPI was used as a nuclear counterstain.
Negative control 1: PBS only.
All lanes : Anti-HDAC2 antibody [Y461] (ab32117) at 1/1000 dilution
Lane 1 : Rat brain tissue homogenate at 20 µg
Lane 2 : Rat brain tissue homogenate at 40 µg
Lane 3 : Rat brain tissue homogenate, P1 nuclear fraction at 20 µg
Lane 4 : Rat brain tissue homogenate, P1 nuclear fraction at 40 µg
Lane 5 : Rat brain tissue homogenate, P2 non-nuclear fraction at 20 µg
Lane 6 : Rat brain tissue homogenate, P2 non-nuclear fraction at 40 µg
All lanes : HRP-conjugated goat anti-rabbit IgG polyclonal at 1/1000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 55 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?
Additional bands at: 35 kDa (possible non-specific binding)
Exposure time: 5 minutes
Anti-HDAC2 antibody [Y461] (ab32117) at 1/2000 dilution + K562 cell lysate
Predicted band size: 55 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?
HDAC2 was immunoprecipitated from 1mg of Hela(Human epithelial cell line from cervix adenocarcinoma)whole cell lysate with ab32117at 1/50 dilution.
Western blot was performed from the immunoprecipitate using ab32117 at 1/1000 dilution.
Anti-Rabbit IgG (HRP),specific to the non-reduced form of IgG, was used as secondary antibody at 1/1000 dilution.
Lane 1: Hela whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Immunohistochemical analysis of HDAC2 expression in paraffin embedded human breast carcinoma tissue section, using 1/250 ab32117.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Overlay histogram showing HeLa cells stained with ab32117 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32117, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
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.
ab32117 被引用在 50 文献中.
- Lin YH et al. HDAC2 (Histone deacetylase 2): A critical factor in environmental enrichment-mediated stroke recovery. J Neurochem N/A:N/A (2020). PubMed: 32415988
- Basta J et al. Pharmacologic inhibition of RGD-binding integrins ameliorates fibrosis and improves function following kidney injury. Physiol Rep 8:e14329 (2020). PubMed: 32281744
- Meng Q et al. Protective effects of histone deacetylase inhibition by Scriptaid on brain injury in neonatal rat models of cerebral ischemia and hypoxia. Int J Clin Exp Pathol 13:179-191 (2020). PubMed: 32211098
- Chung J et al. Protein arginine methyltransferase 5 (PRMT5) promotes survival of lymphoma cells via activation of WNT/ß-catenin and AKT/GSK3ß proliferative signaling. J Biol Chem 294:7692-7710 (2019). PubMed: 30885941
- Oka OB et al. ERp18 regulates activation of ATF6a during unfolded protein response. EMBO J 38:e100990 (2019). PubMed: 31368601