Validated using a knockout cell line
Recombinant
RabMAb

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

Overview

  • Product name
    Anti-USP28 antibody [EPR4249(2)]
    See all USP28 primary antibodies
  • Description
    Rabbit monoclonal [EPR4249(2)] to USP28
  • Tested applications
    Suitable for: WB, Flow Cyt, 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

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

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

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
WB 1/1000 - 1/10000. Detects a band of approximately 140 kDa (predicted molecular weight: 122 kDa).
Flow Cyt Use at an assay dependent concentration.
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



    • Predicted band size : 122 kDa

      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.

    • ab126604 at 1/50 dilution staining USP28 in HeLa cells by Immunofluorescence.
    • 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.

    • 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
      Goat anti-Rabbit HRP at 1/2000 dilution

      Predicted band size : 122 kDa
    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

    References

    This product has been referenced in:
    • 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 . Read more (PubMed: 27432897) »

    See 1 Publication for this product

    Customer reviews and Q&As

    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.

    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 d...

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    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 t...

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