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Anti-hnRNP U/p120 抗体 [3G6] (ab10297)

Reviews (7)Q&A (11)References (26)
Western blot - Anti-hnRNP U/p120 antibody [3G6] (ab10297)
  • Western blot - Anti-hnRNP U/p120 antibody [3G6] (ab10297)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-hnRNP U/p120 antibody [3G6] (ab10297)
  • Flow Cytometry - Anti-hnRNP U/p120 antibody [3G6] (ab10297)

Key features and details

  • Mouse monoclonal [3G6] to hnRNP U/p120
  • Suitable for: WB, IHC-P, Flow Cyt
  • Reacts with: Human
  • Isotype: IgG1

製品の概要

  • 製品名

    Anti-hnRNP U/p120 antibody [3G6]
    hnRNP U/p120 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [3G6] to hnRNP U/p120

  • 由来種

    Mouse

  • アプリケーション

    適用あり: WB, IHC-P, Flow Cytmore details

  • 種交差性

    交差種: Human
    非交差種: Chicken, Lizard

  • 免疫原

    Recombinant full length protein corresponding to Human hnRNP U/p120. RNPs eluted from an oligo (dt) cellulose column.
    Database link: Q00839

  • ポジティブ･コントロール

    • WB: HeLa cytoplasmic and nuclear fraction. HEK-293T whole cell lysate. ICC/IF: U-2 OS cells. IHC-P: Human cerebellum tissue. Flow Cytometry: HeLa cells.

  • 特記事項

    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

製品の特性

  • 製品の状態

    Liquid

  • 保存方法

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

  • バッファー

    Preservative: 0.1% Sodium azide
    Constituent: PBS
  • Concentration information loading...

  • 精製度

    Protein A purified

  • 特記事項（精製）

    Protein A purified from tissue culture supernatant.

  • ポリ/モノ

    モノクローナル

  • クローン名

    3G6

  • ミエローマ

    Sp2/0

  • アイソタイプ

    IgG1

  • 研究分野

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ab10297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
WB (3)
Use at an assay dependent concentration. Detects a band of approximately 97 kDa (predicted molecular weight: 120 kDa).
IHC-P (2)
Use at an assay dependent concentration.
Flow Cyt
Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 
特記事項
WB
Use at an assay dependent concentration. Detects a band of approximately 97 kDa (predicted molecular weight: 120 kDa).
IHC-P
Use at an assay dependent concentration.
Flow Cyt
Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

ターゲット情報

  • 機能

    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.

  • 配列類似性

    Contains 1 B30.2/SPRY domain.
    Contains 1 SAP domain.

  • 翻訳後修飾

    Extensively phosphorylated.
    Arg-733 and Arg-739 are dimethylated, probably to asymmetric dimethylarginine.

  • 細胞内局在

    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

  • 参照データベース

    • 別名

      • 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

    画像

    • Western blot - Anti-hnRNP U/p120 antibody [3G6] (ab10297)
      Western blot - Anti-hnRNP U/p120 antibody [3G6] (ab10297)
      All lanes : Anti-hnRNP U/p120 antibody [3G6] (ab10297) at 1/1000 dilution

      Lane 1 : Cytoplasmic fraction of HeLa (human epithelial cell line from cervix adenocarcinoma) lysate
      Lane 2 : Nuclear fraction of HeLa (human epithelial cell line from cervix adenocarcinoma) lysate

      Predicted band size: 120 kDa
      Observed band size: 120 kDa



      This correlates with the information published in the reference Dreyfuss, G. et. al.
    • Western blot - Anti-hnRNP U/p120 antibody [3G6] (ab10297)
      Western blot - Anti-hnRNP U/p120 antibody [3G6] (ab10297)This image is courtesy of an Abreview submitted by Dr Stoil Dmitrov
      Anti-hnRNP U/p120 antibody [3G6] (ab10297) at 1/5000 dilution + HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 100 µg

      Secondary
      HRP conjugated sheep anti-mouse IgG

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 120 kDa
      Observed band size: 120 kDa
      Additional bands at: 60 kDa, 80 kDa. We are unsure as to the identity of these extra bands.


      Exposure time: 1 minute

      See Abreview

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-hnRNP U/p120 antibody [3G6] (ab10297)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-hnRNP U/p120 antibody [3G6] (ab10297)

      ab10297 (4 µg/ml)) staining hnRNP U/p120 in human cerebellum using an automated system (DAKO Autostainer Plus). Using this protocol there is strong nuclear staining of Purkinje cells neurons.
      Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with a protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

    • Flow Cytometry - Anti-hnRNP U/p120 antibody [3G6] (ab10297)
      Flow Cytometry - Anti-hnRNP U/p120 antibody [3G6] (ab10297)

      Overlay histogram showing HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained with ab10297 (red line). The cells were fixed with 100% methanol (5 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. 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 (ab10297, 1 µg/1x106 cells) for 30 minutes at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 minutes at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 minutes)/permeabilized in 0.1% PBS-Tween used under the same conditions.

    参考文献 (26)

    ab10297 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab10297 は 26 報の論文で使用されています。

    • Zhao L  et al. A Long Non-coding RNA IVRPIE Promotes Host Antiviral Immune Responses Through Regulating Interferon ß1 and ISG Expression. Front Microbiol 11:260 (2020). PubMed: 32153544
    • Zietzer A  et al. The RNA-binding protein hnRNPU regulates the sorting of microRNA-30c-5p into large extracellular vesicles. J Extracell Vesicles 9:1786967 (2020). PubMed: 32944175
    • Sharp JA  et al. Cell division requires RNA eviction from condensing chromosomes. J Cell Biol 219:N/A (2020). PubMed: 33053167
    • Sun WL  et al. Long noncoding RNA OIP5-AS1 targets Wnt-7b to affect glioma progression via modulation of miR-410. Biosci Rep 39:N/A (2019). PubMed: 30498093
    • Alfano L  et al. Depletion of the RNA binding protein HNRNPD impairs homologous recombination by inhibiting DNA-end resection and inducing R-loop accumulation. Nucleic Acids Res N/A:N/A (2019). PubMed: 30799487
    View all Publications for this product

    レビューと Q&A

    Show All レビュー Q&A
    レビューを送る 質問を送る

    1-10 of 18 Abreviews or Q&A

    RIP abreview for Anti-hnRNP U/p120 antibody [3G6]

     Poor
    Abreviews
    Application
    RIP
    Sample
    Mouse Cell lysate - whole cell (Trophoblast Stem Cell)
    Negative control
    IgG
    Immuno-precipitation step
    Protein A/G
    Specification
    Trophoblast Stem Cell
    Detection step
    RIP Seq
    Type
    RIP with cross-linking
    Duration of cross-linking step: 30 minute(s) and 0 second(s)
    Specification of the cross-linking agent: formaldehyde
    Positive control
    Input
    Read More

    Abcam user community

    Verified customer

    投稿 Feb 02 2021

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-hnRNP U/p120 antibody [3G6]

     Excellent
    Abreviews
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Mouse, whole brain sections)
    Antigen retrieval step
    None
    Permeabilization
    Yes - 10% Tween-20
    Specification
    Mouse, whole brain sections
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    投稿 Mar 24 2017

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-hnRNP U/p120 antibody [3G6]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Cerebellum-paraffin embedded)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: pH9
    Specification
    Cerebellum-paraffin embedded
    Blocking step
    Commercial block solution as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    投稿 Dec 29 2016

    Western blot abreview for Anti-hnRNP U/p120 antibody [3G6]

     Excellent
    Abreviews
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Human Embryonic Kidney)
    Gel Running Conditions
    Non-reduced Non-Denaturing (Native) (4-12% Bis-Tris Gel, MES running buffer)
    Loading amount
    15 µg
    Specification
    Human Embryonic Kidney
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    投稿 Apr 01 2014

    Western blot abreview for Anti-hnRNP U antibody [3G6]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Hela cytosolic and nuclear extracts)
    Loading amount
    50 µg
    Specification
    Hela cytosolic and nuclear extracts
    Gel Running Conditions
    Reduced Denaturing (8% gel)
    Blocking step
    Milk as blocking agent for 14 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Read More

    Abcam user community

    Verified customer

    投稿 Nov 12 2009

    Western blot abreview for Anti-hnRNP U/p120 antibody [3G6]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (293T)
    Loading amount
    100 µg
    Specification
    293T
    Blocking step
    Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5%
    Read More

    Dr. Stoil Dmitrov

    Verified customer

    投稿 Apr 09 2007

    Immunocytochemistry/ Immunofluorescence abreview for Anti-hnRNP U/p120 antibody [3G6]

     Good
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (U2OS osteosarcoma line)
    Specification
    U2OS osteosarcoma line
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 1%
    Fixative
    Paraformaldehyde
    Read More

    Dr. Stoil Dmitrov

    Verified customer

    投稿 Apr 06 2007

    Question

    Dear
    I am really appreciate for your understanding, this customer indicated he already bought the other primary antibody from other company,

    therefore, he would like to have a credit note, could you please help him to create a credit note ?

    I deeply appreciate your great assistance

    Best regards,

    Read More
    Answer

    Thank you for confirming those details.

    I'd be happy to arrange for a credit note to be issued. In order to do this, could you please provide the order number on which ab10297 was purchased?

    Many thanks in advance for your cooperation.

    Read More

    Question

    Dear xxxxx
    Thanks for your kindly reply, the questionnaire I attached in this letter, this customer indicated he spend a lot of time to tried on this experiment, and he seems really upset about this state, therefore, could you please help this customer to solve the problem?

    Thanks for your kindly help

    Best regards,

    Read More
    Answer

    Thank you very much for that information.

    I understand the frustration the customer must be feeling and will try to help them as quickly as I can. May I just ask which xxxxxx antibody was used, and which secondary antibody?

    Has the ab10297 been used in a Western blot to ascertain its activity?

    It may unfortunately be that the customer has recieved a faulty vial. If the customer would like, I can provide them with an alternative vial of the antibody, however, we only have the same lot in stock at present. If they would prefer I can provide a replacement with a different antibody such as ab122906. Whilst this antibody has been shown to work in IP, it has not as yet been used with human samples and we would therefore not be able to guarantee that it would work. However, the immunogen used shares 98% homology with the human protein and would therefore be predicted to react.

    Alternatively I can arrange for a credit note to be issued.

    I look forward to hearing how the customer would like to proceed. Many thanks for your cooperation in this case.

    Read More

    Question

    Dear technical support team:

    This adviser already bought ab10297 (Anti-hnRNP U antibody [3G6]) and conducted the IP with MDA-MD231 Whole Cell Lysate sever times, however, the results show no correct band can be detected, and he tried to modified his experiment step many times, but still can’t improve the results, he seems upset about this state, therefore, may I ask for your kindly help to solve this problem as soon as possible?

    He offers some conditions and image in this letter, please check it.
    Lane 1: MDA-MD231 Whole Cell Lysate
    Lanes2-3: used ab10297condcted the IP assay
    Sample: 40ug
    WB: used “santa cruz” antibody (we already measure this antibody is okay!)
    Primary antibody: 1:350 and 1:700
    Secondary antibody: 1:30000
    I appreciate for your kindly help.
    Best regards,

    Read More
    Answer

    Thank you for contacting us and reporting the problems encountered with the anti-hnRNP U antibody [3G6] (ab10297).

    I am sorry to hear youhave beenexperiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

    Thank you for providing the details of the protocol used. In order to understand more fully the protocol used and the samples tested, could you please fill in the questionnaire I have attached to this email?Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary.

    If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.

    I look forward to receiving your reply.

    Read More

    1-10 of 18 Abreviews or Q&A

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