Anti-hnRNP U抗体- ChIP Grade (ab20666)

概述

  • 产品名称Anti-hnRNP U抗体- ChIP Grade
    参阅全部 hnRNP U 一抗
  • 描述
    兔多克隆抗体to hnRNP U - ChIP Grade
  • 经测试应用适用于: ICC/IF, WB, IHC-P, ChIPmore details
  • 种属反应性
    与反应: Mouse, Human
    预测可用于: Rat
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 800 to the C-terminus of Human hnRNP U.

    (Peptide available as ab21993.)

  • 阳性对照
    • This antibody gave a positive signal in HeLa cells, and HepG2 cells, and HeLa whole cell lysate.

性能

应用

Our Abpromise guarantee covers the use of ab20666 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 117 kDa (predicted molecular weight: 90 kDa).
IHC-P 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
ChIP Use at an assay dependent concentration. PubMed: 21235343

靶标

  • 功能Component of the CRD-mediated complex that promotes MYC mRNA stabilization. Binds to pre-mRNA. Has high affinity for scaffold-attached region (SAR) DNA. Binds to double- and single-stranded DNA and RNA.
  • 序列相似性Contains 1 B30.2/SPRY domain.
    Contains 1 SAP domain.
  • 翻译后修饰Extensively phosphorylated.
    Arg-733 and Arg-739 are dimethylated, probably to asymmetric dimethylarginine.
  • 细胞定位Nucleus. Cytoplasm. Cell surface. Localized in cytoplasmic mRNP granules containing untranslated mRNAs. Component of ribonucleosomes. Also found associated with the cell surface.
  • Information by UniProt
  • 数据库链接
  • 别名
    • Heterogeneous nuclear ribonucleoprotein U antibody
    • hnRNP U antibody
    • hnRNP U protein antibody
    • HNRNPU antibody
    • hnRNPU protein antibody
    • HNRPU antibody
    • HNRPU_HUMAN antibody
    • p120 antibody
    • p120 nuclear protein antibody
    • pp120 antibody
    • SAF A antibody
    • SAF-A antibody
    • SAFA antibody
    • Scaffold attachment factor A antibody
    • U21.1 antibody
    see all

Anti-hnRNP U antibody - ChIP Grade 图像

  • ICC/IF image of ab20666 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab20666, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

    Panel A shows localisation of ab20666 to the nuclei, Panel B has the Alexa Fluor® 488 channel removed for comparison.

  • Anti-hnRNP U antibody - ChIP Grade (ab20666) at 1 µg/ml + HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252) at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab7090) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 90 kDa
    Observed band size : 117 kDa (why is the actual band size different from the predicted?)
  • Image courtesy of Human Protein Atlas

    ab20666 staining hnRNP U in paraffin embedded human skin tissue.  The samples were incubated with ab20666 (1/250 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab20666 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines.

    Further results for this antibody can be found at www.proteinatlas.org

  • ab20666 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab20666 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Immunofluorescence analysis of murine embryonic stem cells, staining hnRNP U with ab20666.

    Cells were fixed with paraformaldehyde, permeabilized with 0.25% Triton X-100 and blocked in 0.1% Triton X-100/10% FCS for 20 min. Samples were incubated with primary antibody (1/500) for 2 hours before incubating with an AlexaFluor®488-conjugated goat anti-rabbit IgG (1/500) for 1 hour.

Anti-hnRNP U antibody - ChIP Grade (ab20666)参考文献

This product has been referenced in:
  • Shao R  et al. Coordinate regulation of heterogeneous nuclear ribonucleoprotein dynamics by steroid hormones in the human fallopian tube and endometrium in vivo and in vitro. Am J Physiol Endocrinol Metab 302:E1269-82 (2012). WB, IHC-P ; Human . Read more (PubMed: 22436695) »
  • Hope NR & Murray GI The expression profile of RNA-binding proteins in primary and metastatic colorectal cancer: relationship of heterogeneous nuclear ribonucleoproteins with prognosis. Hum Pathol 42:393-402 (2011). IHC-P ; Human . Read more (PubMed: 21194727) »

See all 4 Publications for this product

Product Wall

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step Heat-inactivated normal donkey serum in 0.05% PBS-T as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Antigen retrieval step None
Sample Mouse Tissue sections (Mouse, whole brain sections, striatum)
Specification Mouse, whole brain sections, striatum
Permeabilization Yes - Tween-20
Fixative Paraformaldehyde
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提交于 Oct 07 2013

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Application Western blot
Loading amount 15 µg
Gel Running Conditions Non-reduced Non-Denaturing (Native) (4-12% Bis-Tris Gel, MES running buffer)
Sample Mouse Tissue lysate - whole (Mouse whole brain tissue lysate, striatum)
Specification Mouse whole brain tissue lysate, striatum
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 Sep 20 2013

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Application Western blot
Sample Human Cell lysate - whole cell (Huh7 cell (Hepatocarcinoma))
Loading amount 100 µg
Specification Huh7 cell (Hepatocarcinoma)
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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提交于 Sep 26 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"