重组Anti-HDAC2抗体[Y461] (ab32117)
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: Human
概述
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产品名称
Anti-HDAC2抗体[Y461]
参阅全部 HDAC2 一抗 -
描述
兔单克隆抗体[Y461] to HDAC2 -
宿主
Rabbit -
特异性
ab32117 recognises HDAC2. -
Tested Applications & Species
Application Species Flow Cyt HumanICC/IF HumanIHC-P HumanIP HumanWB Human -
免疫原
Synthetic peptide within Human HDAC2 aa 450-550 (C terminal). The exact sequence is proprietary.
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阳性对照
- 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.
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常规说明
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.
性能
-
形式
Liquid -
存放说明
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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克隆
单克隆 -
克隆编号
Y461 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- HRP Anti-HDAC2 antibody [Y461] (ab195851)
- Alexa Fluor® 488 Anti-HDAC2 antibody [Y461] (ab196471)
- Alexa Fluor® 647 Anti-HDAC2 antibody [Y461] (ab196518)
- PE Anti-HDAC2 antibody [Y461] (ab210827)
- Anti-HDAC2 antibody [Y461] - BSA and Azide free (ab213700)
- Alexa Fluor® 568 Anti-HDAC2 antibody [Y461] (ab214384)
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
应用
The Abpromise guarantee
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.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
应用 | Species |
---|---|
Flow Cyt |
Human
|
ICC/IF |
Human
|
IHC-P |
Human
|
IP |
Human
|
WB |
Human
|
All applications |
Mouse
Rat
|
应用 | Ab评论 | 说明 |
---|---|---|
WB | (9) |
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. |
|
IP |
1/60.
|
说明 |
---|
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. |
IP
1/60. |
靶标
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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. -
细胞定位
Nucleus. - Information by UniProt
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数据库链接
- 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
see all -
别名
- D10Wsu179e antibody
- HD 2 antibody
- HD2 antibody
see all
图片
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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.
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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).
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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.
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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
Secondary
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
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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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HDAC2 antibody [Y461] (ab32117)
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.
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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.
实验方案
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.
数据表及文件
文献 (50)
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