重组Anti-NF2 / Merlin抗体[EPR2573(2)] (ab109244)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR2573(2)] to NF2 / Merlin
- Suitable for: WB
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
概述
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产品名称
Anti-NF2 / Merlin抗体[EPR2573(2)]
参阅全部 NF2 / Merlin 一抗 -
描述
兔单克隆抗体[EPR2573(2)] to NF2 / Merlin -
宿主
Rabbit -
经测试应用
适用于: WBmore 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
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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 -20°C. Stable for 12 months at -20°C. -
存储溶液
pH: 7.20
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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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR2573(2) -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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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
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab109244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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WB |
1/50000 - 1/200000. Predicted molecular weight: 70 kDa.
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说明 |
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WB
1/50000 - 1/200000. Predicted molecular weight: 70 kDa. |
靶标
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功能
Probable regulator of the Hippo/SWH (Sav/Wts/Hpo) signaling pathway, a signaling pathway that plays a pivotal role in tumor suppression by restricting proliferation and promoting apoptosis. Along with WWC1 can synergistically induce the phosphorylation of LATS1 and LATS2 and can probably function in the regulation of the Hippo/SWH (Sav/Wts/Hpo) signaling pathway. May act as a membrane stabilizing protein. May inhibit PI3 kinase by binding to AGAP2 and impairing its stimulating activity. Suppresses cell proliferation and tumorigenesis by inhibiting the CUL4A-RBX1-DDB1-VprBP/DCAF1 E3 ubiquitin-protein ligase complex. -
组织特异性
Widely expressed. Isoform 1 and isoform 3 are predominant. Isoform 4, isoform 5 and isoform 6 are expressed moderately. Isoform 8 is found at low frequency. Isoform 7, isoform 9 and isoform 10 are not expressed in adult tissues, with the exception of adult retina expressing isoform 10. Isoform 9 is faintly expressed in fetal brain, heart, lung, skeletal muscle and spleen. Fetal thymus expresses isoforms 1, 7, 9 and 10 at similar levels. -
疾病相关
Defects in NF2 are the cause of neurofibromatosis 2 (NF2) [MIM:101000]; also known as central neurofibromatosis. NF2 is a genetic disorder characterized by bilateral vestibular schwannomas (formerly called acoustic neuromas), schwannomas of other cranial and peripheral nerves, meningiomas, and ependymomas. It is inherited in an autosomal dominant fashion with full penetrance. Affected individuals generally develop symptoms of eighth-nerve dysfunction in early adulthood, including deafness and balance disorder. Although the tumors of NF2 are histologically benign, their anatomic location makes management difficult, and patients suffer great morbidity and mortality.
Defects in NF2 are a cause of schwannomatosis (SCHWA) [MIM:162091]; also known as congenital cutaneous neurilemmomatosis. Schwannomas are benign tumors of the peripheral nerve sheath that usually occur singly in otherwise normal individuals. Multiple schwannomas in the same individual suggest an underlying tumor-predisposition syndrome. The most common such syndrome is NF2. The hallmark of NF2 is the development of bilateral vestibular-nerve schwannomas; but two-thirds or more of all NF2-affected individuals develop schwannomas in other locations, and dermal schwannomas may precede vestibular tumors in NF2-affected children. There have been several reports of individuals with multiple schwannomas who do not show evidence of vestibular schwannoma. Clinical report suggests that schwannomatosis is a clinical entity distinct from other forms of neurofibromatosis. -
序列相似性
Contains 1 FERM domain. -
翻译后修饰
Phosphorylation of Ser-518 inhibits nuclear localization by disrupting the intramolecular association of the FERM domain with the C-terminal tail.
Ubiquitinated by the CUL4A-RBX1-DDB1-DCAF1/VprBP E3 ubiquitin-protein ligase complex for ubiquitination and subsequent proteasome-dependent degradation. -
细胞定位
Cytoplasm > perinuclear region. Cytoplasmic granule. Observed in cytoplasmic granules concentrated in a perinuclear location. Isoform 7 is absent from ruffling membranes and filopodia; Cytoplasm > perinuclear region. Cytoplasmic granule. Observed in cytoplasmic granules concentrated in a perinuclear location. Isoform 9 is absent from ruffling membranes and filopodia; Nucleus. Cell projection > filopodium membrane. Cell projection > ruffle membrane. Cytoplasm > perinuclear region. Cytoplasmic granule. Cytoplasm > cytoskeleton. In a fibroblastic cell line, isoform 10 is found homogeneously distributed over the entire cell, with a particularly strong staining in ruffling membranes and filopodia and Cell projection > filopodium membrane. Cell projection > ruffle membrane. Nucleus. In a fibroblastic cell line, isoform 1 is found homogeneously distributed over the entire cell, with a particularly strong staining in ruffling membranes and filopodia. Colocalizes with MPP1 in non-myelin-forming Schwann cells. Binds with VPRBP in the nucleus. The intramolecular association of the FERM domain with the C-terminal tail promotes nuclear accumulation. The unphosphorylated form accumulates predominantly in the nucleus while the phosphorylated form is largely confined to the non-nuclear fractions. - Information by UniProt
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数据库链接
- Entrez Gene: 4771 Human
- Entrez Gene: 18016 Mouse
- Entrez Gene: 25744 Rat
- Omim: 607379 Human
- SwissProt: P35240 Human
- SwissProt: P46662 Mouse
- SwissProt: Q63648 Rat
- Unigene: 187898 Human
see all -
别名
- ACN antibody
- BANF antibody
- Bilateral acoustic neuroma antibody
see all
图片
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All lanes : Anti-NF2 / Merlin antibody [EPR2573(2)] (ab109244) at 1/50000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : NF2 knockout A549 cell lysate
Lane 3 : SH-SY5Y cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 70 kDa
Observed band size: 73 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-NF2 / Merlin antibody [EPR2573(2)] staining at 1/50000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab109244 was shown to bind specifically to NF2 / Merlin. A band was observed at 73 kDa in wild-type A549 cell lysates with no signal observed at this size in NF2 knockout cell line. To generate this image, wild-type and NF2 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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All lanes : Anti-NF2 / Merlin antibody [EPR2573(2)] (ab109244) at 1/50000 dilution
Lane 1 : Wild-type HepG2 cell lysate
Lane 2 : NF2 knockout HepG2 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 70 kDa
Observed band size: 74 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-NF2 / Merlin antibody [EPR2573(2)] staining at 1/50000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab109244 was shown to bind specifically to NF2 / Merlin. A band was observed at 74 kDa in wild-type HepG2 cell lysates with no signal observed at this size in NF2 knockout cell line ab277859 (knockout cell lysate ab283826). To generate this image, wild-type and NF2 knockout HepG2 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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All lanes : Anti-NF2 / Merlin antibody [EPR2573(2)] (ab109244) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : NF2 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 70 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 - ab109244 observed at 60 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab109244 was shown to react with NF2 in wild-type HeLa cells in western blot with loss of signal observed in NF2 knockout cell line ab261796 (NF2 knockout cell lysate ab257179). Wild-type and NF2 knockout HeLa cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab109244 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 hour at room temperature before imaging.
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Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: NF2 knockout HAP1 whole cell lysate (20 µg)Lanes 1 - 2: Merged signal (red and green). Green - ab109244 observed at 70 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab109244 was shown to specifically react with NF2 in wild type cells as signal was lost in NF2 knockout cells. Wild-type and NF2 knockout samples were subjected to SDS-PAGE. Ab109244 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 50000 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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All lanes : Anti-NF2 / Merlin antibody [EPR2573(2)] (ab109244) at 1/50000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : MCF7 cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : PC3 cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 70 kDa
数据表及文件
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SDS download
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Datasheet download
文献 (6)
ab109244 被引用在 6 文献中.
- Tian Q et al. RICH1 inhibits breast cancer stem cell traits through activating kinases cascade of Hippo signaling by competing with Merlin for binding to Amot-p80. Cell Death Dis 13:71 (2022). PubMed: 35064101
- Jia Z et al. A novel NF2 splicing mutant causes neurofibromatosis type 2 via liquid-liquid phase separation with large tumor suppressor and Hippo pathway. iScience 25:105275 (2022). PubMed: 36300003
- Vichas A et al. Integrative oncogene-dependency mapping identifies RIT1 vulnerabilities and synergies in lung cancer. Nat Commun 12:4789 (2021). PubMed: 34373451
- Sikorski K et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). PubMed: 30377371
- Guan Y et al. Knockdown of miR-572 suppresses cell proliferation and promotes apoptosis in renal cell carcinoma cells by targeting the NF2/Hippo signaling pathway. Int J Clin Exp Pathol 11:5705-5714 (2018). PubMed: 31949656
- Pelossof R et al. Prediction of potent shRNAs with a sequential classification algorithm. Nat Biotechnol 35:350-353 (2017). PubMed: 28263295