Overview

  • Product name

    Anti-USP28 antibody [EPR4249(2)] - BSA and Azide free
    See all USP28 primary antibodies
  • Description

    Rabbit monoclonal [EPR4249(2)] to USP28 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt, ICC/IF, WBmore 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

    Ab225537 is the carrier-free version of ab126604. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab225537 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

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

Properties

Applications

Our Abpromise guarantee covers the use of ab225537 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 Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

ICC/IF Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 140 kDa (predicted molecular weight: 122 kDa).
  • 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

    • This WB data was generated using the same anti-USP28 antibody clone [EPR4249(2)] in a different buffer formulation (cat# 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.

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126604).

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

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126604).

    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126604).

    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

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