重组Anti-PABPN1抗体[EP3000Y] (ab75855)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP3000Y] to PABPN1
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-P
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-PABPN1抗体[EP3000Y]
参阅全部 PABPN1 一抗 -
描述
兔单克隆抗体[EP3000Y] to PABPN1 -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-Pmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide within Human PABPN1 aa 1-100 (N terminal). The exact sequence is proprietary.
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阳性对照
- WB: Raw264.7, MCF-7, 293T, Mouse spleen, Rat brain and HeLa whole cell lysate (ab150035). ICC/IF: MCF-7 cells. Flow Cyt (intra): MCF-7 cells. IHC-P: Squamous cell cervical carcinoma tissue, human bladder carcinoma, mouse kidney, rat kidney.
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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. Stable for 12 months at -20°C. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EP3000Y -
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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Positive Controls
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab75855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) |
1/40.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1:80. |
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ICC/IF |
1/100 - 1/250.
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WB | (1) |
1/1000 - 1/10000. Predicted molecular weight: 33 kDa.
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IP |
1/30.
Fur unpurified use at 1:50. |
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IHC-P |
1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. |
说明 |
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Flow Cyt (Intra)
1/40. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1:80. |
ICC/IF
1/100 - 1/250. |
WB
1/1000 - 1/10000. Predicted molecular weight: 33 kDa. |
IP
1/30. Fur unpurified use at 1:50. |
IHC-P
1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols. |
靶标
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功能
Involved in the 3'-end formation of mRNA precursors (pre-mRNA) by the addition of a poly(A) tail of 200-250 nt to the upstream cleavage product. Stimulates poly(A) polymerase (PAPOLA) conferring processivity on the poly(A) tail elongation reaction and controls also the poly(A) tail length. Increases the affinity of poly(A) polymerase for RNA. Is also present at various stages of mRNA metabolism including nucleocytoplasmic trafficking and nonsense-mediated decay (NMD) of mRNA. Cooperates with SKIP to synergistically activate E-box-mediated transcription through MYOD1 and may regulate the expression of muscle-specific genes. Binds to poly(A) and to poly(G) with high affinity. May protect the poly(A) tail from degradation. -
组织特异性
Ubiquitous. -
疾病相关
Defects in PABPN1 are the cause of oculopharyngeal muscular dystrophy (OPMD) [MIM:164300]. OPMD is a form of late-onset slowly progressive myopathy characterized by eyelid ptosis, dysphagia and, sometimes by other cranial and limb-muscle involvement. -
序列相似性
Contains 1 RRM (RNA recognition motif) domain. -
结构域
The RRM domain is essential for specific adenine bases recognition in the poly(A) tail but not sufficient for poly(A) binding. -
翻译后修饰
Arginine dimethylation is asymmetric and involves PRMT1 and PRMT3. It does not influence the RNA binding properties. -
细胞定位
Nucleus. Cytoplasm. Localized in cytoplasmic mRNP granules containing untranslated mRNAs. Shuttles between the nucleus and the cytoplasm but predominantly found in the nucleus. Its nuclear import may involve the nucleocytoplasmic transport receptor transportin and a RAN-GTP-sensitive import mechanism. Is exported to the cytoplasm by a carrier-mediated pathway that is independent of mRNA traffic. Nucleus; nuclear speckle. Colocalizes with SKIP and poly(A) RNA in nuclear speckles. - Information by UniProt
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数据库链接
- Entrez Gene: 8106 Human
- Entrez Gene: 54196 Mouse
- Entrez Gene: 116697 Rat
- Omim: 602279 Human
- SwissProt: Q86U42 Human
- SwissProt: Q8CCS6 Mouse
- Unigene: 707712 Human
- Unigene: 7723 Mouse
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别名
- Nuclear poly(A)-binding protein 1 antibody
- OPMD antibody
- PAB2 antibody
see all
图片
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Anti-PABPN1 antibody [EP3000Y] (ab75855) at 1/2000 dilution (purified) + Rat brain lysates at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 33 kDaBlocking and diluting buffer: 5% NFDM/TBST
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Intracellular Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling PABPN1 with purified ab75855 at 1/40 dilution (10 ug/ml) (red). Cells were fixed with 80% Methanol and permeabilized with 0.1% Tween-20. A Goat anti rabbit IgG (Alexa Fluorr®488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling PABPN1 with purified ab75855 at 1:100 dilution (4.1μg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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ab75855 (purified) at 1:30 dilution (5ug) immunoprecipitating PABPN1 in MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate.
Lane 1 (input): MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate 10ug
Lane 2 (+): ab75855 & MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab75855 in MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:10,000 dilution. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue sections labeling PABPN1 with Purified ab75855 at 1:1000 dilution (0.41 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling PABPN1 with Purified ab75855 at 1:1000 dilution (0.41 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human bladder carcinoma tissue sections labeling PABPN1 with Purified ab75855 at 1:1000 dilution (0.41 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
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Immunocytochemistry/Immunofluorescence analysis of MCF-7 cells labelling PABPN1 with unpurified ab75855 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody.
Control: PBS only.
Nuclear counter stain: DAPI. -
All lanes : Anti-PABPN1 antibody [EP3000Y] (ab75855) at 1/2000 dilution (purified)
Lane 1 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates
Lane 2 : Mouse spleen lysates
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 33 kDaBlocking and diluting buffer: 5% NFDM/TBST
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All lanes : Anti-PABPN1 antibody [EP3000Y] (ab75855) at 1/2000 dilution (purified)
Lane 1 : 293T (Human embryonic kidney epithelial cell) whole cell lysates
Lane 2 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 33 kDa
Observed band size: 49 kDa why is the actual band size different from the predicted?Blocking and diluting buffer: 5% NFDM/TBST
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Lanes 1-2 : Anti-PABPN1 antibody [EP3000Y] (ab75855) at 1/200000 dilution (unpurified)
Lane 3 : Anti-PABPN1 antibody [EP3000Y] (ab75855) at 1/1000000 dilution (unpurified)
Lane 1 : Raw264.7 cell lysate
Lane 2 : MCF-7 cell lysate
Lane 3 : HeLa cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP-conjugated goat anti-rabbit IgG at 1/1000 dilution
Predicted band size: 33 kDa
Observed band size: 49 kDa why is the actual band size different from the predicted? -
Overlay histogram showing MCF-7 cells stained with unpurified ab75855 (red line). The cells were fixed with 80% 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 (ab75855, 1µg/1x106 cells) 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 IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
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Unpurified ab75855, at 1/100 dilution, staining PABPN1 in squamous cell cervical carcinoma, by Immunohistochemistry using formalin-fixed, paraffin-embedded tissue.
Perform heat mediated antigen retrieval 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
文献 (39)
ab75855 被引用在 39 文献中.
- Kajjo S et al. PABP prevents the untimely decay of select mRNA populations in human cells. EMBO J 41:e108650 (2022). PubMed: 35156721
- Nicholson-Shaw AL et al. Nuclear and cytoplasmic poly(A) binding proteins (PABPs) favor distinct transcripts and isoforms. Nucleic Acids Res 50:4685-4702 (2022). PubMed: 35438785
- Roth F et al. Assessment of PABPN1 nuclear inclusions on a large cohort of patients and in a human xenograft model of oculopharyngeal muscular dystrophy. Acta Neuropathol 144:1157-1170 (2022). PubMed: 36197469
- Qi JZ et al. Functions of long non-coding RNA LNC11649 in non-small cell lung cancer cells as a reprocessed form of MALAT1. Neoplasma 69:1322-1337 (2022). PubMed: 36264775
- Booy EP et al. The noncoding RNA BC200 associates with polysomes to positively regulate mRNA translation in tumor cells. J Biol Chem 296:100036 (2021). PubMed: 33410401