Anti-Ubiquitin 抗体 [Ubi-1] (ab7254)

Reviews (6)Q&A (21)References (51)

Key features and details

  • Mouse monoclonal [Ubi-1] to Ubiquitin
  • Suitable for: IHC-P, WB
  • Reacts with: Mouse, Chicken, Cow, Human, Arabidopsis thaliana, Caenorhabditis elegans, Drosophila melanogaster
  • Isotype: IgG1

製品の概要

  • 製品名

    Anti-Ubiquitin antibody [Ubi-1]
    Ubiquitin 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [Ubi-1] to Ubiquitin

  • 由来種

    Mouse

  • アプリケーション

    適用あり: IHC-P, WBmore details
    適用なし: IP

  • 種交差性

    交差種: Mouse, Chicken, Cow, Human, Arabidopsis thaliana, Caenorhabditis elegans, Drosophila melanogaster

  • 免疫原

    Full length protein corresponding to Ubiquitin conjugated to Keyhole Limpet Haemocyanin (KLH). Mouse monoclonal [Ubi-1] to Ubiquitin was raised against purified ubiquitin conjugated with glutaraldehyde to keyhole limpet hemocyanin.

  • ポジティブ･コントロール

    • H9C2 cells IHC-P:FFPE mouse brain alzheimer

  • 特記事項

    This product was changed from ascites to tissue culture supernatant on 17th May 2016. The following lots are from ascites and are still in stock as of 17th May 2016: GR159581-2, GR159581-12, GR159581-13. Lot numbers higher than GR159581-13 will be from tissue culture supernatant. Please note that the dilutions may need to be adjusted accordingly.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab7254 in the following tested applications.

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

アプリケーション Abreviews 特記事項
IHC-P Use a concentration of 1 - 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/100 - 1/5000.
  • 追加情報
    Is unsuitable for IP.

    • ターゲット情報

    • 関連性

      Function: Ubiquitin exists either covalently attached to another protein, or free (unanchored). When covalently bound, it is conjugated to target proteins via an isopeptide bond either as a monomer (monoubiquitin), a polymer linked via different Lys residues of the ubiquitin (polyubiquitin chains) or a linear polymer linked via the initiator Met of the ubiquitin (linear polyubiquitin chains). Polyubiquitin chains, when attached to a target protein, have different functions depending on the Lys residue of the ubiquitin that is linked: Lys-6-linked may be involved in DNA repair; Lys-11-linked is involved in ERAD (endoplasmic reticulum-associated degradation) and in cell-cycle regulation; Lys-29-linked is involved in lysosomal degradation; Lys-33-linked is involved in kinase modification; Lys-48-linked is involved in protein degradation via the proteasome; Lys-63-linked is involved in endocytosis, DNA-damage responses as well as in signaling processes leading to activation of the transcription factor NF-kappa-B. Linear polymer chains formed via attachment by the initiator Met lead to cell signaling. Ubiquitin is usually conjugated to Lys residues of target proteins, however, in rare cases, conjugation to Cys or Ser residues has been observed. When polyubiquitin is free (unanchored-polyubiquitin), it also has distinct roles, such as in activation of protein kinases, and in signaling. Similarity: Belongs to the ubiquitin family. Contains 3 ubiquitin-like domains.

    • 細胞内局在

      Cell Membrane, Cytoplasmic and Nuclear

    • 参照データベース

      see all

    • 別名

      • Epididymis secretory protein Li 50 antibody
      • FLJ25987 antibody
      • HEL S 50 antibody
      • MGC8385 antibody
      • Polyubiquitin B antibody
      • RPS 27A antibody
      • RPS27A antibody
      • UBA 52 antibody
      • UBA 80 antibody
      • UBA52 antibody
      • UBA80 antibody
      • UBB antibody
      • UBB_HUMAN antibody
      • UBC antibody
      • UBCEP 1 antibody
      • UBCEP 2 antibody
      • UBCEP1 antibody
      • UBCEP2 antibody
      • Ubiquitin antibody
      • Ubiquitin B antibody
      see all

    画像

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin antibody [Ubi-1] (ab7254)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin antibody [Ubi-1] (ab7254)Image from Ohmi K et al., Plos One. 2011;6(11):e27461. Fig 7.; doi: 10.1371/journal.pone.0027461 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

      Immunohistochemical staining of ubiquitin in a 7 months-old MPS IIIA mouse brain (PFA fixed) using ab7254.

      Sections, 40 µm thick, were cut sagittally on a vibratome. Slices were permeabilized in 1% triton X-100/PBS for 1 hour at room temperature. The samples were placed on vectabond-coated glass slides and dried overnight. They were then rehydrated in PBS and washed in ice-cold methanol for 10 min.

      The slices were washed in 0.3% H2O2 in methanol at room temperature and incubated sequentially with 0.5% normal donkey serum, ab7254 and biotin-labeled F(ab')2 donkey secondary antibody in Tris-buffered saline (pH 7.5) containing 2% BSA and 0.02% Tween 20. The signal was detected with an ABC kit, visualized with diaminobenzidine and counterstained with hematoxylin.

    • Western blot - Anti-Ubiquitin antibody [Ubi-1] (ab7254)
      Western blot - Anti-Ubiquitin antibody [Ubi-1] (ab7254)
      All lanes : Anti-Ubiquitin antibody [Ubi-1] (ab7254) at 1/100 dilution

      Lane 1 : HeLa cell lysate
      Lane 2 : HeLa cell lysate with MG132
      Lane 3 : Mouse brain tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Anti mouse IgG at 1/10000 dilution
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin antibody [Ubi-1] (ab7254)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin antibody [Ubi-1] (ab7254)

      IHC image of ab7254 staining in mouse alzheimer brain formalin fixed paraffin embedded tissue section, using MOM detection kit, ab127055. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was incubated with ab7254, 5µg/ml, for 15 mins at room temperature.  DAB was used as the chromogen (ab103723). The section was 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin antibody [Ubi-1] (ab7254)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin antibody [Ubi-1] (ab7254)

      Human normal placenta (ab29745). Staining is localised in the cytoplasm and in the nuclei. Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control. Sections were stained using an automated system DAKO Autostainer Plus, at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers citrate pH6.1 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for mouse 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 we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

    • Western blot - Anti-Ubiquitin antibody [Ubi-1] (ab7254)
      Western blot - Anti-Ubiquitin antibody [Ubi-1] (ab7254)Image courtesy of an anonymous Abreview.
      HeLa cells were co-transfected with a plasmid expressing a target protein together with Ubi expressing vector for 24 hours and either left untreated (Contr) or were treated with 10 µM MG-132 for 6 hours (+MG132). Then the protein of interest was pulled down using Flag agarose beads and and probed with ab7254 at a 1/2000 dilution. The secondary used was an Alexa-Fluor 680 conjugated goat anti-mouse polyclonal used at a 1/10000 dilution. The protein is known to be degraded through proteasome.

      See Abreview

    参考文献 (51)

    ab7254 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。

    ab7254 は 51 報の論文で使用されています。

    • Shin D  et al. Regulation of Phosphoribosyl-Linked Serine Ubiquitination by Deubiquitinases DupA and DupB. Mol Cell 77:164-179.e6 (2020). PubMed: 31732457
    • Lo Piccolo L  et al. The lncRNA hsr? regulates arginine dimethylation of human FUS to cause its proteasomal degradation in Drosophila. J Cell Sci 132:N/A (2019). PubMed: 31519807
    • Sharma A  et al. UVR8 disrupts stabilisation of PIF5 by COP1 to inhibit plant stem elongation in sunlight. Nat Commun 10:4417 (2019). PubMed: 31562307
    • Yuan Y  et al. Mitochondrial ROS-induced lysosomal dysfunction impairs autophagic flux and contributes to M1 macrophage polarization in a diabetic condition. Clin Sci (Lond) 133:1759-1777 (2019). PubMed: 31383716
    • Schmidt H  et al. IL-13 Impairs Tight Junctions in Airway Epithelia. Int J Mol Sci 20:N/A (2019). PubMed: 31262043
    View all Publications for this product

    レビューと Q&A

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    1-10 of 27 Abreviews or Q&A

    Western blot abreview for Anti-Ubiquitin antibody [Ubi-1]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Cardiomyocytes)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    25 µg
    Treatment
    Starvation
    Specification
    Cardiomyocytes
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    投稿 Nov 11 2016

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Ubiquitin [Ubi-1] antibody

     Good
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: RT°C
    Sample
    Human Cell (Human primary fibroblasts)
    Specification
    Human primary fibroblasts
    Permeabilization
    Yes - 0.2% Triton-X100
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    投稿 Mar 26 2014

    Question

    What is the species of origin for the ubiquitin used as immunogen for this antibody?

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    Answer

    Thank you for your enquiry.

    The immunogen was purified bovine ubiquitin isolated from erythrocytes and cross-linked into an insoluble complex by glutaraldehyde. Ubiquitin is one of the most conserved proteins and the sequence is virtually identical across all vertebrates, and identical in mammals.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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    Question

    Please book ab7254 .

    Regards

    Read More
    Answer

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement of ab7254 on your order ######. This has Abcam order reference #####.

    To check the status of the order please contact our Customer Service team and reference this number.

    Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

    I wish you the customer best of luck with their research.

    Read More

    Question

    Here is the order no ######
    Regards

    Read More
    Answer

    Thank you for taking the time to complete the questionnaire and provide the order number. I am sorry to hear the customer has had difficulty obtaining satisfactory results from this antibody.

    The details you have kindly provided will provide us with vital information for our monitoring of product quality

    I appreciate the time the customer has spent in the laboratory and understand your concerns. It is regrettable the results have not been succesful. Reviewing the details, I am sorry there are no further tips to provide on this occasion to help improve the results. I can suggest you have regrettably recieved a bad vial.

    I apologise for the inconvenience and am pleased to offer you a free of charge replacement or credit note in compensation.

    In addition, I can suggest you may like to consider the following suggestions which I hope will be helpful:

    1. I notice from the order records that when we were first contacted, this was just over the 6 month guarantee period. I am pleased to provide an exception in this case as it is only a few weeks over. However, please note that we encourage customers to contact us as soon as possible if they encounter any difficulties, and within the 6 month guarantee period.

    2. Has the quality of the sample been assessed with a loading control?

    Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

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    Question

    I have purchased 2 vials of ab7254 recently. They are both the same lot number. The first one worked well but the second one is detecting very little or no ubiquitin. I have used an antibody against the ubiquitinated protein to prove it is there and I see ub forms of this protein with that antibody.

    Read More
    Answer

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab7254.

    To check the status of the order please contact our Customer Service team and reference this number.

    Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

    I wish you the best of luck with your research.

    Read More

    Question

    I used to anti-ubiquitin antibodies with the same free floating sections with antigen pH 6 retrieval buffer using mouse brain. One antibody worked great (ab7254) and another ab7780 didn't stain at all.

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    Answer

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab7254.

    To check the status of the order please contact our Customer Service team and reference this number.

    Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

    I wish you the best of luck with your research.

    Read More

    Question

    I’m in AB4587 and don’t see a link to MSDS.  I see the Data sheet link to the right, but no link to msds. Could you look at this page and tell me where it is?   https://www.abcam.com/Alzheimer-brain-human-temporal-lobe-tissue-slides-ab4587.html    

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    Answer

    Thanks for your email. ab4587 does not have any required MSDS information, so a link will not be present on this datasheet. For example, ab7254 does have a link to MSDS information. I hope this helps. Please contact me again if you have any further questions. 

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    Question

        I’m looking for MSDS for the items below. B-Amyloid Control Slides (5 Slides/PK) ABCAM AB4587 BCL2 Antibody ABCAM AB32124 BETA Amyloid ABCAM   BLIMP-1 ABCAM AB81961 FOXP3 ANTIBODY (236A/E7) ABCAM WIHC031 GCET1 ABCAM AB68889 GPNMB Antibody (Monoclonal) (100ug) ABCAM AB56584 Human Milk Fat Globule 1 Antibody  (500 uL) ABCAM AB17787 IMP-3 antibody ABCAM AB82044 Mesothelioma Positive Controls ABCAM AB4437 Oct1 Antibody (500uL) ABCAM AB15112 SMOOTHELIN ANTIBODY ABCAM   TFE-3 ABCAM AB70008 Ubiquitin ABCAM AB7254      

    Read More
    Answer

    Thanks for your enquiry. If required, MSDS information is located via one or more 'MSDS' links near the top left corner of the online datasheet. This link is just below the 'Datasheet PDF' link present on all datasheets. I hope this is helpful. Please contact me again if you have any further questions.

    Read More

    Western blot abreview for Anti-Ubiquitin antibody [Ubi-1]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa cells)
    Loading amount
    100 µg
    Specification
    HeLa cells
    Treatment
    10 µM MG-132 for 6hrs
    Gel Running Conditions
    Reduced Denaturing (10% Tris-Gly gel)
    Blocking step
    LI-COR® Odyssey® Blocking Buffer as blocking agent for 45 minute(s) · Concentration: 50% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    投稿 Dec 13 2011

    1-10 of 27 Abreviews or Q&A

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