重组Anti-RING2 / RING1B / RNF2抗体[EPR12245] (ab181140)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR12245] to RING2 / RING1B / RNF2
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
概述
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产品名称
Anti-RING2 / RING1B / RNF2抗体[EPR12245]
参阅全部 RING2 / RING1B / RNF2 一抗 -
描述
兔单克隆抗体[EPR12245] to RING2 / RING1B / RNF2 -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details -
种属反应性
与反应: Mouse, Human
预测可用于: Rat -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- HeLa, 293, Jurkat and HepG2 whole cell lysate (ab7900); Human placenta tissue; HepG2 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
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 59% PBS, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR12245 -
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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Positive Controls
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab181140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) |
Use at an assay dependent concentration.
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WB |
1/1000 - 1/10000. Detects a band of approximately 41 kDa (predicted molecular weight: 38 kDa).
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IHC-P | (3) |
1/250 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/100 - 1/250.
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说明 |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
WB
1/1000 - 1/10000. Detects a band of approximately 41 kDa (predicted molecular weight: 38 kDa). |
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/100 - 1/250. |
靶标
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功能
E3 ubiquitin-protein ligase that mediates monoubiquitination of 'Lys-119' of histone H2A, thereby playing a central role in histone code and gene regulation. H2A 'Lys-119' ubiquitination gives a specific tag for epigenetic transcriptional repression and participates in X chromosome inactivation of female mammals. May be involved in the initiation of both imprinted and random X inactivation. Essential component of the Polycomb group (PcG) multiprotein PRC1 complex, a complex required to maintain the transcriptionally repressive state of many genes, including Hox genes, throughout development. PcG PRC1 complex act via chromatin remodeling and modification of histones, rendering chromatin heritably changed in its expressibility. E3 ubiquitin-protein ligase activity is enhanced by BMI1/PCGF4. Acts as the main E3 ubiquitin ligase on histone H2A of the PRC1 complex, while RING1 may rather act as a modulator of RNF2/RING2 activity. -
通路
Protein modification; protein ubiquitination. -
序列相似性
Contains 1 RING-type zinc finger. -
翻译后修饰
Polyubiquitinated in the presence of UBE2D3 (in vitro).
Monoubiquitinated, by auto-ubiquitination. -
细胞定位
Nucleus. Chromosome. Enriched on inactive X chromosome (Xi) in female trophoblast stem (TS) cells as well as differentiating embryonic stem (ES) cells. The enrichment on Xi is transient during TS and ES cell differentiation. The association with Xi is mitotically stable in non-differentiated TS cells. - Information by UniProt
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数据库链接
- Entrez Gene: 6045 Human
- Entrez Gene: 19821 Mouse
- Entrez Gene: 304850 Rat
- Omim: 608985 Human
- SwissProt: Q99496 Human
- SwissProt: Q9CQJ4 Mouse
- SwissProt: Q4KLY4 Rat
- Unigene: 591490 Human
see all -
别名
- BAP 1 antibody
- BAP1 antibody
- DING antibody
see all
图片
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All lanes : Anti-RING2 / RING1B / RNF2 antibody [EPR12245] (ab181140) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : RNF2 CRISPR/Cas9 edited HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 38 kDa
Observed band size: 42 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab181140 observed at 42 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab181140 was shown to react with RING2 / RING1B / RNF2 in wild-type HeLa cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab264845 (CRISPR/Cas9 edited cell lysate ab257640) lane below 42kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and RNF2 CRISPR/Cas9 edited HeLa 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. ab181140 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated 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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Immunohistochemical analysis of paraffin-embedded Human placenta tissue labeling RING2 / RING1B / RNF2 with ab181140 at 1/500 dilution followed by pre-diluted HRP-conjugated secondary antibody and counter-stained with Hematoxylin.
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: RING2 / RING1B / RNF2 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: Jurkat cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab181140 observed at 42 kDa. Red - loading control, ab7291, observed at 52 kDa.
ab181140 was shown to recognize RING2 / RING1B / RNF2 when RING2 / RING1B / RNF2 knockout samples were used, along with additional cross-reactive bands. Wild-type and RING2 / RING1B / RNF2 knockout samples were subjected to SDS-PAGE. ab181140 at a dilution of 1/1000 and ab7291 (loading control to alpha Tubulin) at a dilution of 1/10,000 were incubated overnight at 4°C. 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 1/10,000 dilution for 1 h at room temperature before imaging. -
Intracellular Flow Cytometry analysis of HepG2 (human hepatocellular carcinoma) cells labeling RING2 / RING1B / RNF2 with purified ab181140 at 1/70 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
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All lanes : Anti-RING2 / RING1B / RNF2 antibody [EPR12245] (ab181140) at 1/10000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : 293 cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : HepG2 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution
Predicted band size: 38 kDa -
Immunofluorescent analysis of HepG2 cells (paraformaldehyde-fixed, 4%) labeling RING2 / RING1B / RNF2 with ab181140 at 1/250 dilution followed by Goat anti rabbit IgG (Alexa Fluor® 555) secondary at 1/200 dilution and counter-stained with DAPI (blue).
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 (6)
ab181140 被引用在 6 文献中.
- Li L et al. The Spatiotemporal Expression of Ring1B in the Postnatal Mouse Cochlea. J Int Adv Otol 18:297-301 (2022). PubMed: 35894525
- Gong Z et al. Exogenous melatonin prevents type 1 diabetes mellitus-induced bone loss, probably by inhibiting senescence. Osteoporos Int 33:453-466 (2022). PubMed: 34519833
- Chen L et al. PHC1 maintains pluripotency by organizing genome-wide chromatin interactions of the Nanog locus. Nat Commun 12:2829 (2021). PubMed: 33990559
- Maat H et al. The USP7-TRIM27 axis mediates non-canonical PRC1.1 function and is a druggable target in leukemia. iScience 24:102435 (2021). PubMed: 34113809
- Liu H et al. G-protein-coupled receptor GPR17 inhibits glioma development by increasing polycomb repressive complex 1-mediated ROS production. Cell Death Dis 12:610 (2021). PubMed: 34120140
- Azkanaz M et al. Protein quality control in the nucleolus safeguards recovery of epigenetic regulators after heat shock. Elife 8:N/A (2019). PubMed: 31199242