Anti-U2AF65抗体(ab37530)
Key features and details
- Rabbit polyclonal to U2AF65
- Suitable for: IP, IHC-P, ChIP, ICC/IF, WB
- Reacts with: Mouse, Human, Zebrafish
- Isotype: IgG
选择批间可重复性更高的重组抗体
- 研究可靠 —— 各批次间结果一致且可重复
- 长期批量供应 —— 采用重组技术,可实现快速生产
- 首次实验即可成功 —— 经过大量验证确认了特异性
- 符合伦理标准 —— 产品不含动物成分
概述
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产品名称
Anti-U2AF65抗体
参阅全部 U2AF65 一抗 -
描述
兔多克隆抗体to U2AF65 -
宿主
Rabbit -
经测试应用
适用于: IP, IHC-P, ChIP, ICC/IF, WBmore details -
种属反应性
与反应: Mouse, Human, Zebrafish
预测可用于: Cow, Xenopus laevis -
免疫原
Synthetic peptide corresponding to Human U2AF65 aa 200-300.
(Peptide available asab37529) -
常规说明
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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 or -80°C. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading...
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纯度
Immunogen affinity purified -
克隆
多克隆 -
同种型
IgG -
研究领域
相关产品
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Compatible Secondaries
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Isotype control
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab37530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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IP |
Use at an assay dependent concentration.
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IHC-P |
Use a concentration of 5 µg/ml.
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ChIP |
Use at an assay dependent concentration. PubMed: 27252533
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ICC/IF | (1) |
Use a concentration of 1 µg/ml.
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WB | (1) |
1/250. Detects a band of approximately 65 kDa (predicted molecular weight: 53 kDa).
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说明 |
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IP
Use at an assay dependent concentration. |
IHC-P
Use a concentration of 5 µg/ml. |
ChIP
Use at an assay dependent concentration. PubMed: 27252533 |
ICC/IF
Use a concentration of 1 µg/ml. |
WB
1/250. Detects a band of approximately 65 kDa (predicted molecular weight: 53 kDa). |
靶标
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功能
Necessary for the splicing of pre-mRNA. Induces cardiac troponin-T (TNNT2) pre-mRNA exon inclusion in muscle. Regulates the TNNT2 exon 5 inclusion through competition with MBNL1. Binds preferentially to a single-stranded structure within the polypyrimidine tract of TNNT2 intron 4 during spliceosome assembly. Required for the export of mRNA out of the nucleus, even if the mRNA is encoded by an intron-less gene. Represses the splicing of MAPT/Tau exon 10. -
序列相似性
Belongs to the splicing factor SR family.
Contains 3 RRM (RNA recognition motif) domains. -
翻译后修饰
Lysyl-hydroxylation at Lys-15 and Lys-276 affects the mRNA splicing activity of the protein, leading to regulate some, but not all, alternative splicing events. -
细胞定位
Nucleus. - Information by UniProt
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数据库链接
- Entrez Gene: 11338 Human
- Entrez Gene: 22185 Mouse
- Entrez Gene: 380287 Xenopus laevis
- Omim: 191318 Human
- SwissProt: P26368 Human
- SwissProt: P26369 Mouse
- Unigene: 528007 Human
- Unigene: 360389 Mouse
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别名
- hU2AF(65) antibody
- hU2AF65 antibody
- Splicing factor U2AF 65 kDa subunit antibody
see all
图片
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ab37530 staining U2AF65 in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab37530 at 5µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue). Also suitable in cells fixed with 4% paraformaldehyde (10 min).Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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All lanes : Anti-U2AF65 antibody (ab37530) at 1/250 dilution
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 :Jurkat whole cell lysate (ab7899)
Lane 3 : A-431 whole cell lysate (ab7909)
Lane 4 : HEK-293 whole cell lysate (ab7902)
Lane 5 : Hep G2 whole cell lysate (ab7900)
Lane 6 : MCF-7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 7 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
Lane 8 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Rabbit IgG secondary antibody (ab28446) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 53 kDa
Observed band size: 65 kDa why is the actual band size different from the predicted?Although the predicted band size is 53kDa based on Swiss-prot data, a band of 65kDa has been previously observed. J Biol Chem. 2004 Nov 26;279(48):49773-9 (PMID: 15377657)
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-U2AF65 antibody (ab37530)IHC image of ab37530 staining in Human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab37530, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. -
All lanes : Anti-U2AF65 antibody (ab37530) at 1/250 dilution
Lane 1 : Marker
Lane 2 : Zebrafish brain homogenate at 20 µg
Lane 3 : Zebrafish heart homogenate at 10 µg
Lane 4 : Zebrafish liver homogenate at 10 µg
Lane 5 : Zebrafish skeletal muscle homogenate at 10 µg
Lane 6 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
Secondary
All lanes : Goat polyclonal to Rabbit IgG – H&L – Pre-Adsorbed (HRP) at 1/6000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 53 kDa
Observed band size: 53 kDa
Exposure time: 5 minutes -
U2AF65 was immunoprecipitated using 0.5mg SHSY5Y whole cell extract, 5µg of Rabbit polyclonal to U2AF65 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, SHSY5Y whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab37530.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 65kDa: U2AF65.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (22)
ab37530 被引用在 22 文献中.
- Isaac R et al. A seven-transmembrane protein-TM7SF3, resides in nuclear speckles and regulates alternative splicing. iScience 25:105270 (2022). PubMed: 36304109
- Zhu Q et al. LINC00173 promotes Wilms' tumor progression through MGAT1-mediated MUC3A N-glycosylation. Cell Cycle 21:1795-1810 (2022). PubMed: 35491865
- Ma N et al. RETRACTED: circTulp4 functions in Alzheimer’s disease pathogenesis by regulating its parental gene, Tulp4. Mol Ther 29:2167-2181 (2021). PubMed: 33578037
- Wang L et al. A novel tumour suppressor protein encoded by circMAPK14 inhibits progression and metastasis of colorectal cancer by competitively binding to MKK6. Clin Transl Med 11:e613 (2021). PubMed: 34709743
- Xu J et al. LncRNA BBOX1-AS1 upregulates HOXC6 expression through miR-361-3p and HuR to drive cervical cancer progression. Cell Prolif 53:e12823 (2020). PubMed: 32515533