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遺伝子ノックアウト細胞株 検証済リコンビナントRabMAb

Anti-USP22 抗体 [EPR4352(2)] (ab109435)

Reviews (1)Q&A (2)References (1)
Western blot - Anti-USP22 antibody [EPR4352(2)] (ab109435)
  • Western blot - Anti-USP22 antibody [EPR4352(2)] (ab109435)
  • Western blot - Anti-USP22 antibody [EPR4352(2)] (ab109435)
  • Flow Cytometry (Intracellular) - Anti-USP22 antibody [EPR4352(2)] (ab109435)
  • OI-RD Scanning - Anti-USP22 antibody [EPR4352(2)] (ab109435)
  • Anti-USP22 antibody [EPR4352(2)] (ab109435)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4352(2)] to USP22
  • Suitable for: Flow Cyt (Intra), WB
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

製品の概要

  • 製品名

    Anti-USP22 antibody [EPR4352(2)]
    USP22 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [EPR4352(2)] to USP22

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: Flow Cyt (Intra), WBmore details
    適用なし: ICC/IF or IHC-P

  • 種交差性

    交差種: Human

  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • ポジティブ･コントロール

    • WB: HeLa, HAP1, human placenta and HT-1376 lysates

  • 特記事項

    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
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

製品の特性

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ab109435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
Flow Cyt (Intra)
1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.  
WB
1/1000 - 1/5000. Predicted molecular weight: 60 kDa.
特記事項
Flow Cyt (Intra)
1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.  
WB
1/1000 - 1/5000. Predicted molecular weight: 60 kDa.
追加情報
Is unsuitable for ICC/IF or IHC-P.

ターゲット情報

  • 機能

    Histone deubiquitinating component of the transcription regulatory histone acetylation (HAT) complex SAGA. Catalyzes the deubiquitination of both histones H2A and H2B, thereby acting as a coactivator. Recruited to specific gene promoters by activators such as MYC, where it is required for transcription. Required for nuclear receptor-mediated transactivation and cell cycle progression.

  • 組織特異性

    Moderately expressed in various tissues including heart and skeletal muscle, and weakly expressed in lung and liver.

  • 配列類似性

    Belongs to the peptidase C19 family. UBP8 subfamily.
    Contains 1 UBP-type zinc finger.

  • 細胞内局在

    Nucleus.
  • Information by UniProt

  • 参照データベース

    • 別名

      • Deubiquitinating enzyme 22 antibody
      • KIAA1063 antibody
      • Ubiquitin carboxyl terminal hydrolase 22 antibody
      • Ubiquitin carboxyl-terminal hydrolase 22 antibody
      • Ubiquitin specific peptidase 22 antibody
      • Ubiquitin specific peptidase 3 like antibody
      • Ubiquitin specific processing protease 22 antibody
      • Ubiquitin specific protease 22 antibody
      • Ubiquitin thioesterase 22 antibody
      • Ubiquitin thiolesterase 22 antibody
      • Ubiquitin-specific-processing protease 22 antibody
      • UBP22_HUMAN antibody
      • USP 22 antibody
      • Usp22 antibody
      • USP3L antibody
      see all

    画像

    • Western blot - Anti-USP22 antibody [EPR4352(2)] (ab109435)
      Western blot - Anti-USP22 antibody [EPR4352(2)] (ab109435)
      All lanes : Anti-USP22 antibody [EPR4352(2)] (ab109435) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : USP22 knockout HeLa cell lysate

      Performed under reducing conditions.

      Predicted band size: 60 kDa
      Observed band size: 59 kDa why is the actual band size different from the predicted?



      False colour image of Western blot: Anti-USP22 antibody [EPR4352(2)] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab109435 was shown to bind specifically to USP22. A band was observed at 59 kDa in wild-type HeLa cell lysates with no signal observed at this size in usp22 knockout cell line ab264888 (knockout cell lysate ab257789). To generate this image, wild-type and usp22 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

    • Western blot - Anti-USP22 antibody [EPR4352(2)] (ab109435)
      Western blot - Anti-USP22 antibody [EPR4352(2)] (ab109435)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: USP22 knockout HAP1 cell lysate (20 µg)
      Lane 3: HeLa cell lysate (20 µg)
      Lane 4: HepG2 cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab109435 observed at 60 kDa. Red - loading control, ab8245, observed at 37 kDa.


      Ab109435 was shown to specifically react with USP22 in wild-type cells along with additional cross-reactive bands. The band was not seen in USP22 knockout HAP1 cells. Wild-type and USP22 knockout samples were subjected to SDS-PAGE. ab109435 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

    • Western blot - Anti-USP22 antibody [EPR4352(2)] (ab109435)
      Western blot - Anti-USP22 antibody [EPR4352(2)] (ab109435)
      All lanes : Anti-USP22 antibody [EPR4352(2)] (ab109435) at 1/1000 dilution

      Lane 1 : HeLa cell lysate
      Lane 2 : Human placenta lysate
      Lane 3 : HT-1376 lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 60 kDa

    • Flow Cytometry (Intracellular) - Anti-USP22 antibody [EPR4352(2)] (ab109435)
      Flow Cytometry (Intracellular) - Anti-USP22 antibody [EPR4352(2)] (ab109435)

      Intracellular flow cytometric analysis of permeabilized HeLa cells using 1/10 ab109435 (red) or a rabbit IgG (negative) (green). 

       

    • OI-RD Scanning - Anti-USP22 antibody [EPR4352(2)] (ab109435)
      OI-RD Scanning - Anti-USP22 antibody [EPR4352(2)] (ab109435)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • Anti-USP22 antibody [EPR4352(2)] (ab109435)
      Anti-USP22 antibody [EPR4352(2)] (ab109435)

    参考文献 (1)

    ab109435 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。

    ab109435 は 1 報の論文で使用されています。

    • Liu Z  et al. USP22 regulates the formation and function of placental vasculature during the development of fetal growth restriction. Placenta 111:19-25 (2021). PubMed: 34130183

    レビューと Q&A

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    レビューを送る 質問を送る

    1-3 of 3 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-USP22 antibody [EPR4352(2)]

     Good
    Abreviews
    This product is known to not work in this application or species.
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Specification
    HeLa
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.5% Triton X100
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    DR. Kirk Mcmanus

    Verified customer

    投稿 Sep 16 2011

    Question

    Thank you. I appreciate your help. This is the reason that we always purchase antibodies from abcam.

    I would like to have two different antibodies as replacement. These are:anti-GFP (ab290) and anti-H3 (ab1791). Please let me know if you have a reference# for the replacement.

    Thanks!

    Read More
    Answer

    As requested, I have issued free of charge replacements.


    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

    This is a follow-up on my earlier inquiry from a while back (see below). I'm reattaching the data that we have which shows the antibody is not performing. To answer the questions in your reply:

    (1) For the RFP antibody, on the blot, we expected to see mCherry at around 25kDa, the same size as EGFP. What was loaded was bacterial lysate that expresses mCherry, which is visibly red. So we know that the protein is present. The "none" lane is bacterial lysate without any expression plasmid present. We also checked that your immunogen sequence is present in the coding sequence of mCherry.

    (2) The blot is blocked with 5% milk in TBST. As you can see, the GFP antibody performed well.

    (3) For the Usp22 antibody, what was loaded were HeLa cells lysed in 1% SDS. Lysate was sonicated before loaded on the gel. 5 mM bME was present in the sample loading buffer. So it is under denaturing conditions. No protease inhibitor is needed since we lyse cells directly in SDS, which will quickly kill proteases as well.

    Please let me know your thoughts. We are confident that these antibodies are not performing. We are long-time customers and hope abcam honor your guarantee. Thanks.

    Read More
    Answer

    Thank you for following up your earlier complaints about these two antibodies, ab28664 and ab109435, with more details of your protocols and samples.

    I will be happy to replace them. We have not received other complaints about them but I suggest trying two different antibodies.

    For the RFP antibody, I suggest a mouse monoclonal, ab109809:

    Click here (or use the following: https://www.abcam.com/RFP-antibody-RFPH8-ab109809.html).

    We unfortunately do not have an alternative to anti-USP22 ab109435, or even a different lot. If you would like an antibody against a different target instead, I can send that, or a credit or refund.

    I look forward to your reply.

    Read More

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