Anti-nmt55 / p54nrb 抗体 [EPR5270] (ab133574)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR5270] to nmt55 / p54nrb
- Suitable for: WB, ICC/IF, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-nmt55 / p54nrb antibody [EPR5270]
nmt55 / p54nrb 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR5270] to nmt55 / p54nrb -
由来種
Rabbit -
アプリケーション
適用あり: WB, ICC/IF, IHC-Pmore details -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Synthetic peptide within Human nmt55/ p54nrb (C terminal). The exact sequence is proprietary.
Database link: Q15233 -
ポジティブ・コントロール
- WB: HEK293T, PC3, MOLT4, HeLa, MCF7, A673, RAW 264.7, PC12 and NIH/3T3 cell lysates. IHC-P: Human kidney and bladder carcinoma tissues.
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特記事項
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.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. -
解離定数(KD 値)
KD = 2.20 x 10 -12 M Learn more about KD -
バッファー
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine) -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPR5270 -
アイソタイプ
IgG -
研究分野
関連製品
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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
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab133574の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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WB |
1/1000 - 1/10000. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa).
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ICC/IF |
1/1500.
For unpurified use at 1/250 - 1/500. |
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IHC-P |
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. Use a HRP/AP polymerized secondary antibody. |
特記事項 |
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WB
1/1000 - 1/10000. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa). |
ICC/IF
1/1500. For unpurified use at 1/250 - 1/500. |
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols. Use a HRP/AP polymerized secondary antibody. |
ターゲット情報
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機能
DNA- and RNA binding protein, involved in several nuclear processes. Binds the conventional octamer sequence in double stranded DNA. Also binds single-stranded DNA and RNA at a site independent of the duplex site (By similarity). Involved in pre-mRNA splicing, probably as an heterodimer with SFPQ. Interacts with U5 snRNA, probably by binding to a purine-rich sequence located on the 3' side of U5 snRNA stem 1b. The SFPQ-NONO heteromer associated with MATR3 may play a role in nuclear retention of defective RNAs. The SFPQ-NONO heteromer may be involved in DNA unwinding by modulating the function of topoisomerase I/TOP1. The SFPQ-NONO heteromer may be involved in DNA nonhomologous end joining (NHEJ) required for double-strand break repair and V(D)J recombination and may stabilize paired DNA ends. In vitro, the complex strongly stimulates DNA end joining, binds directly to the DNA substrates and cooperates with the Ku70/G22P1-Ku80/XRCC5 (Ku) dimer to establish a functional preligation complex. Nono is involved in transcriptional regulation. The SFPQ-NONO-NR5A1 complex binds to the CYP17 promoter and regulates basal and cAMP-dependent transcriptional avtivity. NONO binds to an enhancer element in long terminal repeats of endogenous intracisternal A particles (IAPs) and activates transcription. -
組織特異性
Heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas. Also found in a number of breast tumor cell lines. -
関連疾患
Note=A chromosomal aberration involving NONO may be a cause of papillary renal cell carcinoma (PRCC). Translocation t(X;X)(p11.2;q13.1) with TFE3. -
配列類似性
Contains 2 RRM (RNA recognition motif) domains. -
翻訳後修飾
The N-terminus is blocked. -
細胞内局在
Nucleus. - Information by UniProt
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参照データベース
- Entrez Gene: 4841 Human
- Entrez Gene: 53610 Mouse
- Entrez Gene: 317259 Rat
- Omim: 300084 Human
- SwissProt: Q15233 Human
- SwissProt: Q99K48 Mouse
- SwissProt: Q5FVM4 Rat
- Unigene: 533282 Human
see all -
別名
- 52 kDa subunit antibody
- 54 kDa nuclear RNA and DNA binding protein antibody
- 54 kDa nuclear RNA- and DNA-binding protein antibody
see all
画像
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All lanes : Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : NONO knockout HEK293T cell lysate
Lane 3 : MOLT-4 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 54 kDa
Observed band size: 63 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab133574 observed at 63 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab133574 Anti-nmt55 / p54nrb antibody [EPR5270] was shown to specifically react with nmt55 / p54nrb in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266244 (knockout cell lysate ab257160) was used. Wild-type and nmt55 / p54nrb knockout samples were subjected to SDS-PAGE. ab133574 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution 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 10000 dilution for 1 hour at room temperature before imaging.
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Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574) at 1/10000 dilution (purified) + PC-12 cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 54 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST. -
Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574) at 1/10000 dilution (purified) + RAW264.7 cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 54 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST. -
All lanes : Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574) at 1/10000 dilution (purified)
Lane 1 : Molt-4 cell lysate
Lane 2 : A673 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 54 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST. -
All lanes : Anti-nmt55 / p54nrb antibody [EPR5270] (ab133574) at 1/1000 dilution (unpurified)
Lane 1 : PC3 cell lysate
Lane 2 : MOLT4 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : MCF7 cell lysate
Lane 5 : A673 cell lysate
Lane 6 : RAW 264.7 cell lysate
Lane 7 : PC12 cell lysate
Lane 8 : NIH/3T3 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Standard HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 54 kDa
Observed band size: 54 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue labelling nmt55/p54nrb with purified ab133574 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue labelling nmt55/p54nrb with purified ab133574 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate tissue labelling nmt55/p54nrb with purified ab133574 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human bladder carcinoma tissue labelling nmt55 / p54nrb using unpurified ab133574 at a 1/250 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human kidney tissue labelling nmt55 /p54nrb using unpurified ab133574 at a 1/250 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunocytochemistry/Immunofluorescence analysis of Molt-4 (human acute lymphoblastic leukemia) cells labelling nmt55/p54nrb with purified ab133574 at 1/1500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (6)
ab133574 は 6 報の論文で使用されています。
- Wang S et al. Long non-coding RNA SNHG1 suppresses cell migration and invasion and upregulates SOCS2 in human gastric carcinoma. Biochem Biophys Rep 27:101052 (2021). PubMed: 34179518
- Pan YY et al. RNA-binding protein SFPQ cooperates with HDAC1 to suppress CD40 transcription in pulmonary adventitial fibroblasts. Cell Biol Int 44:166-176 (2020). PubMed: 31393052
- Barra J et al. Integrator restrains paraspeckles assembly by promoting isoform switching of the lncRNA NEAT1. Sci Adv 6:eaaz9072 (2020). PubMed: 32923585
- Shen W et al. Acute hepatotoxicity of 2' fluoro-modified 5-10-5 gapmer phosphorothioate oligonucleotides in mice correlates with intracellular protein binding and the loss of DBHS proteins. Nucleic Acids Res 46:2204-2217 (2018). PubMed: 29390093
- Hupalowska A et al. CARM1 and Paraspeckles Regulate Pre-implantation Mouse Embryo Development. Cell 175:1902-1916.e13 (2018). PubMed: 30550788
- Shadle SC et al. DUX4-induced dsRNA and MYC mRNA stabilization activate apoptotic pathways in human cell models of facioscapulohumeral dystrophy. PLoS Genet 13:e1006658 (2017). ICC/IF ; Human . PubMed: 28273136