重组Anti-HDAC2抗体[EPR20117] (ab219053)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20117] to HDAC2
- Suitable for: Flow Cyt (Intra), ChIC/CUT&RUN-seq, IHC-P, ICC/IF, IP, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-HDAC2抗体[EPR20117]
参阅全部 HDAC2 一抗 -
描述
兔单克隆抗体[EPR20117] to HDAC2 -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), ChIC/CUT&RUN-seq, IHC-P, ICC/IF, IP, WBmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: His-tagged human HDAC2 recombinant protein (aa339-488);HeLa,SH-SY5Y,HEK-293,PC-12,NIH/3T3 whole cell lysates; Human fetal brain,fetal heart and fetal kidney lysates; Mouse brain and heart lysates; Rat heart,brain and spleen lysates IHC-P: Human testis,tonsil,prostate hyperplasia,prostate cancer,breast cancer and synovial sarcoma tissues; mouse colon tissue and rat spleen tissue ICC/IF: HEK-293 and NIH/3T3 cells Flow Cyt (intra): NIH/3T3 cells IP: HeLa cell lysate ChIC/CUT&RUN-Seq: K-562 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
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形式
Liquid -
存放说明
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. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR20117 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab219053于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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Flow Cyt (Intra) |
1/500.
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ChIC/CUT&RUN-seq |
Use at an assay dependent concentration.
5µg |
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IHC-P |
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/1000.
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IP |
1/30.
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WB |
1/1000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).
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说明 |
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Flow Cyt (Intra)
1/500. |
ChIC/CUT&RUN-seq
Use at an assay dependent concentration. 5µg |
IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/1000. |
IP
1/30. |
WB
1/1000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa). |
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
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别名
- D10Wsu179e antibody
- HD 2 antibody
- HD2 antibody
see all
图片
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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 ab172730 is also shown.
Additional screenshots of mapped reads can be downloaded here.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
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All lanes : Anti-HDAC2 antibody [EPR20117] (ab219053) at 1/1000 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 - ab219053 observed at 55 kDa. Red - 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 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. ab219053 and 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 (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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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) (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 (ab172730) instead of ab219053 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
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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) (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 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) (ab150077) at 1/1000 dilution.
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All lanes : Anti-HDAC2 antibody [EPR20117] (ab219053) at 1/1000 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 - ab219053 observed at 60 kDa. Red - 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 ab266590 (knockout cell lysate ab256939) was used. Wild-type and HDAC2 knockout samples were subjected to SDS-PAGE. ab219053 and 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 (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 [EPR20117] (ab219053) at 1/1000 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 - ab219053 observed at 60 kDa. Red - 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 ab266588 (knockout cell lysate ab256937) was used. Wild-type and HDAC2 knockout samples were subjected to SDS-PAGE. ab219053 and 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 (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 [EPR20117] (ab219053) at 1/1000 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 - ab219053 observed at 55 kDa. Red - 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 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 ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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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.
-
All lanes : Anti-HDAC2 antibody [EPR20117] (ab219053) at 1/1000 dilution
Lane 1 : His-tagged human HDAC2 recombinant protein (aa339-488)
Lane 2 : His-tagged human HDAC1 recombinant protein (aa1-482)
Lysates/proteins at 0.01 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 55 kDa
Observed band size: 22 kDa why is the actual band size different from the predicted?
Exposure time: 1 secondBlocking/Dilution buffer: 5% NFDM/TBST.
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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 (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.
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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) (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 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) (ab150077) at 1/1000 dilution.
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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.
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All lanes : Anti-HDAC2 antibody [EPR20117] (ab219053) at 1/5000 dilution
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate
Lane 3 : HEK-293 (Human epithelial cell line from embryonic kidney) 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
Predicted band size: 55 kDa
Observed band size: 55 kDa
Exposure time: 10 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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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.
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All lanes : Anti-HDAC2 antibody [EPR20117] (ab219053) at 1/5000 dilution
Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution
Predicted band size: 55 kDa
Observed band size: 55 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1: 3 minutes; Lane 2: 15 seconds; Lane 3: 2 seconds.
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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.
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Lanes 1-5 : Anti-HDAC2 antibody [EPR20117] (ab219053) at 1/1000 dilution
Lanes 6-7 : Anti-HDAC2 antibody [EPR20117] (ab219053) at 1/5000 dilution
Lane 1 : Mouse brain lysate
Lane 2 : Mouse heart lysate
Lane 3 : Rat heart lysate
Lane 4 : Rat brain lysate
Lane 5 : Rat spleen lysate
Lane 6 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 7 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 55 kDa
Observed band size: 55 kDaBlocking/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.
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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.
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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.
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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.
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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.
实验方案
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (1)
ab219053 被引用在 1 文献中.
- Yang X et al. FKBP3 Induces Human Immunodeficiency Virus Type 1 Latency by Recruiting Histone Deacetylase 1/2 to the Viral Long Terminal Repeat. mBio 12:e0079521 (2021). PubMed: 34281390