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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-USP28 antibody [EPR4249(2)] (ab126604)

Submit a review Q&A (3)References (8)
Western blot - Anti-USP28 antibody [EPR4249(2)] (ab126604)
  • Immunocytochemistry/ Immunofluorescence - Anti-USP28 antibody [EPR4249(2)] (ab126604)
  • Flow Cytometry (Intracellular) - Anti-USP28 antibody [EPR4249(2)] (ab126604)
  • Western blot - Anti-USP28 antibody [EPR4249(2)] (ab126604)
  • OI-RD Scanning - Anti-USP28 antibody [EPR4249(2)] (ab126604)
  • Anti-USP28 antibody [EPR4249(2)] (ab126604)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4249(2)] to USP28
  • Suitable for: Flow Cyt (Intra), WB, ICC/IF
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free PE

Overview

  • Product name

    Anti-USP28 antibody [EPR4249(2)]
    See all USP28 primary antibodies

  • Description

    Rabbit monoclonal [EPR4249(2)] to USP28

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, ICC/IFmore details
    Unsuitable for: IHC-P or IP

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide, corresponding to residues in Human USP28.

  • Positive control

    • 293T, HeLa, HT1376, SW480 and A431 cell lysates; Human heart tissue lysate; HeLa cells.

  • 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.

    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 ab126604 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)
Use at an assay dependent concentration.
WB
1/1000 - 1/10000. Detects a band of approximately 140 kDa (predicted molecular weight: 122 kDa).
ICC/IF
1/50 - 1/100.
Notes
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB
1/1000 - 1/10000. Detects a band of approximately 140 kDa (predicted molecular weight: 122 kDa).
ICC/IF
1/50 - 1/100.
Application notes
Is unsuitable for IHC-P or IP.

Target

  • Function

    Deubiquitinase involved in DNA damage response checkpoint and MYC proto-oncogene stability. Involved in DNA damage induced apoptosis by specifically deubiquitinating proteins of the DNA damage pathway such as CLSPN. Also involved in G2 DNA damage checkpoint, by deubiquitinating CLSPN, and preventing its degradation by the anaphase promoting complex/cyclosome (APC/C). In contrast, it does not deubiquitinate PLK1. Specifically deubiquitinates MYC in the nucleoplasm, leading to prevent MYC degradation by the proteasome: acts by specifically interacting with isoform 1 of FBXW7 (FBW7alpha) in the nucleoplasm and counteracting ubiquitination of MYC by the SCF(FBW7) complex. In contrast, it does not interact with isoform 4 of FBXW7 (FBW7gamma) in the nucleolus, allowing MYC degradation and explaining the selective MYC degradation in the nucleolus.

  • Sequence similarities

    Belongs to the peptidase C19 family. USP28 subfamily.
    Contains 1 UIM (ubiquitin-interacting motif) repeat.

  • Post-translational
    modifications

    Degradaded upon nickel ion level or hypoxia exposure.
    Phosphorylated upon DNA damage at Ser-67 and Ser-714, by ATM or ATR.

  • Cellular localization

    Nucleus > nucleoplasm.
  • Information by UniProt

  • Database links

    • Alternative names

      • Deubiquitinating enzyme 28 antibody
      • KIAA1515 antibody
      • Ubiquitin carboxyl terminal hydrolase 28 antibody
      • Ubiquitin carboxyl-terminal hydrolase 28 antibody
      • ubiquitin carboxyl-terminal hydrolase 28 variant 1 antibody
      • Ubiquitin specific peptidase 28 antibody
      • Ubiquitin specific processing protease 28 antibody
      • Ubiquitin specific protease 28 antibody
      • Ubiquitin thioesterase 28 antibody
      • Ubiquitin thiolesterase 28 antibody
      • Ubiquitin-specific-processing protease 28 antibody
      • UBP28_HUMAN antibody
      • USP 28 antibody
      • USP28 antibody
      • USP28 protein antibody
      see all

    Images

    • Western blot - Anti-USP28 antibody [EPR4249(2)] (ab126604)
      Western blot - Anti-USP28 antibody [EPR4249(2)] (ab126604)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: USP28 knockout HAP1 cell lysate (20 µg)
      Lane 3: HeLa cell lysate (20 µg)
      Lane 4: A431 cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab126604 observed at 128 kDa. Red - loading control, ab8245, observed at 37 kDa.
      ab126604 was shown to recognize USP28 when USP28 knockout samples were used, along with additional cross-reactive bands. Wild-type and USP28 knockout samples were subjected to SDS-PAGE. ab126604 and ab8245 (loading control to GAPDH) were diluted 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 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-USP28 antibody [EPR4249(2)] (ab126604)
      Immunocytochemistry/ Immunofluorescence - Anti-USP28 antibody [EPR4249(2)] (ab126604)
      ab126604 at 1/50 dilution staining USP28 in HeLa cells by Immunofluorescence.
    • Flow Cytometry (Intracellular) - Anti-USP28 antibody [EPR4249(2)] (ab126604)
      Flow Cytometry (Intracellular) - Anti-USP28 antibody [EPR4249(2)] (ab126604)

      Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling USP28 with unpurified ab126604 at 1/200 dilution (1ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) (ab172730) was used as the isotype control, Cell without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control. 

       

       

    • Western blot - Anti-USP28 antibody [EPR4249(2)] (ab126604)
      Western blot - Anti-USP28 antibody [EPR4249(2)] (ab126604)
      All lanes : Anti-USP28 antibody [EPR4249(2)] (ab126604) at 1/1000 dilution

      Lane 1 : 293T cell lysate
      Lane 2 : HeLa cell lysate
      Lane 3 : HT1376 cell lysate
      Lane 4 : Human heart tissue lysate
      Lane 5 : SW480 cell lysate
      Lane 6 : A431 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit HRP at 1/2000 dilution

      Predicted band size: 122 kDa

    • OI-RD Scanning - Anti-USP28 antibody [EPR4249(2)] (ab126604)
      OI-RD Scanning - Anti-USP28 antibody [EPR4249(2)] (ab126604)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • Anti-USP28 antibody [EPR4249(2)] (ab126604)
      Anti-USP28 antibody [EPR4249(2)] (ab126604)

    References (8)

    Publishing research using ab126604? Please let us know so that we can cite the reference in this datasheet.

    ab126604 has been referenced in 8 publications.

    • Ni T  et al. PTBP1 drives c-Myc-dependent gastric cancer progression and stemness. Br J Cancer 128:1005-1018 (2023). PubMed: 36635500
    • Zhang A  et al. USP33 deubiquitinates and stabilizes HIF-2alpha to promote hypoxia response in glioma stem cells. EMBO J 41:e109187 (2022). PubMed: 35191554
    • Wang H  et al. USP28 and USP25 are downregulated by Vismodegib in vitro and in colorectal cancer cell lines. FEBS J 288:1325-1342 (2021). PubMed: 32578360
    • Chen L  et al. USP28 facilitates pancreatic cancer progression through activation of Wnt/β-catenin pathway via stabilising FOXM1. Cell Death Dis 12:887 (2021). PubMed: 34584067
    • Ruiz EJ  et al. USP28 deletion and small-molecule inhibition destabilizes c-MYC and elicits regression of squamous cell lung carcinoma. Elife 10:N/A (2021). PubMed: 34636321
    • Liu J  et al. Arginine methylation-dependent LSD1 stability promotes invasion and metastasis of breast cancer. EMBO Rep 21:e48597 (2020). PubMed: 31833203
    • Müller I  et al. Cancer Cells Employ Nuclear Caspase-8 to Overcome the p53-Dependent G2/M Checkpoint through Cleavage of USP28. Mol Cell 77:970-984.e7 (2020). PubMed: 31982308
    • Meitinger F  et al. 53BP1 and USP28 mediate p53 activation and G1 arrest after centrosome loss or extended mitotic duration. J Cell Biol 214:155-66 (2016). Human . PubMed: 27432897

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-3 of 3 Abreviews or Q&A

    Question

    I have a vial of this USP28 Ab (ab126604). Two specific questions: 1. Is this antibody tested for ELISA application? 2. What's the region of protein the antibody is recognizing (i.e. region of immunogen sequence)? I need to have an idea the recognition site may not be blocked by other antibodies.

    Read More
    Answer

    The USP28 antibody ab126604 has not, to our knowledge, been tested in ELISA. It was raised against a sequence near the C terminus of the protein, between amino acids 120 - 160 of accession Q96RU2, http://www.uniprot.org/uniprot/Q96RU2.

    Read More

    Question

    can yuo give me an offer for ab137438?
    thank you

    Read More
    Answer

    DISCOUNT CODE: XXXXXXX
    Expiration date: 06-26-2013
    Value: $349

    I am very pleased to hear you would like to accept our offer and test ab137438 in Pig (Sus Scrofa). This code will give you $349 off your next order before the expiration date. To redeem this offer, please submit an Abreview for Pig and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
    For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.


    The terms and conditions applicable to this offer can be found here: www.abcam.com/abtrial.

    Read More

    Question

    I would like to test the antibody Abcam ID: ab126604 (anti-USP28 antibodies) in pig tissues. Can you give me an abtrial discount code?
    thank you

    Read More
    Answer

    Thank you for contacting us.

    The immunogen sequence alignment of ab126604 with Pig protein (http://www.uniprot.org/uniprot/F1SM65) is just 20% which means there are minimal changes that this antibody would cross react. I am sorry I am unable to offer you 100% Abtrial discount code.

    The following antibody immunogen show high similarity e.g. ab137438, ab70896 (http://www.ebi.ac.uk/Tools/services/web_clustalw2/toolresult.ebi?tool=clustalw2&jobId=clustalw2-I20130227-113829-0962-37379653-oy)

    I can provide Abtrial code for these ones if you are interested.

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

    Read More

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