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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-USP22 antibody [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

Overview

  • Product name

    Anti-USP22 antibody [EPR4352(2)]
    See all USP22 primary antibodies

  • Description

    Rabbit monoclonal [EPR4352(2)] to USP22

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), WBmore details
    Unsuitable for: ICC/IF or IHC-P

  • Species reactivity

    Reacts with: Human

  • Immunogen

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

  • Positive control

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

  • General notes

    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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

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

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab109435 in the following tested applications.

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

Application Abreviews Notes
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.
Notes
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.
Application notes
Is unsuitable for ICC/IF or IHC-P.

Target

  • Function

    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.

  • Tissue specificity

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

  • Sequence similarities

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

  • Cellular localization

    Nucleus.
  • Information by UniProt

  • Database links

    • Alternative names

      • 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

    Images

    • 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)

    References (1)

    Publishing research using ab109435? Please let us know so that we can cite the reference in this datasheet.

    ab109435 has been referenced in 1 publication.

    • 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

    Customer reviews and Q&As

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    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

    Submitted 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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