Overview

  • Product name

  • Description

    Rabbit polyclonal to UBPY/USP8
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human UBPY/USP8 aa 400-450. The exact sequence is proprietary. NP_005145.3
    Database link: P40818

  • Positive control

    • WB: HeLa and HEK-293T whole cell lysates. IP: HeLa whole cell lysate. IHC-P: Human breast carcinoma tissue.

Properties

Applications

Our Abpromise guarantee covers the use of ab245641 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
IP Use at 2-5 µg/mg of lysate.
WB 1/2000 - 1/10000. Predicted molecular weight: 128 kDa.
IHC-P 1/200 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • Function

    Hydrolase that can remove conjugated ubiquitin from proteins and therefore plays an important regulatory role at the level of protein turnover by preventing degradation. Converts both 'Lys-48' an 'Lys-63'-linked ubiquitin chains. Catalytic activity is enhanced in the M phase. Involved in cell proliferation. Required to enter into S phase in response to serum stimulation. May regulate T-cell anergy mediated by RNF128 via the formation of a complex containing RNF128 and OTUB1. Probably regulates the stability of STAM2 and RASGRF1. Regulates endosomal ubiquitin dynamics, cargo sorting, membrane traffic at early endosomes, and maintenance of ESCRT-0 stability. The level of protein ubiquitination on endosomes is essential for maintaining the morphology of the organelle. Deubiquitinates EPS15 and controles tyrosine kinase stability. Removes conjugated ubiquitin from EGFR thus regulating EGFR degradation and downstream MAPK signaling. Involved in acrosome biogenesis through interaction with the spermatid ESCRT-0 complex and microtubules. Deubiquitinates BIRC6/bruce and KIF23/MKLP1.
  • Sequence similarities

    Belongs to the peptidase C19 family.
    Contains 1 MIT domain.
    Contains 1 rhodanese domain.
    Contains 1 USP domain.
  • Domain

    The MIT domain is required for endosomal localization, CHMP1B-binding, maintenance of ESCRT-0 stability and EGFR degradation.
    The rhodanese domain is sufficient for RNF41-binding.
  • Post-translational
    modifications

    Phosphorylation of Ser-718 is essential for interaction with YWHAE and for cytosol localization. Undergoes dephosphorylation at Ser-718 in the M phase. Tyrosine-phosphorylated in its N-terminal half in an EGFR-dependent manner.
    Ubiquitinated. Inactive form is mostly monoubiquitinated, but polyubiquitination happens too. Ubiquitination is increased in EGF-stimulated cells. Ubiquitination of active form is undetectable, suggesting a possibility that USP8 deubiquitinates itself, thereby regulating its own function.
  • Cellular localization

    Cytoplasm. Nucleus. Endosome membrane. Cell membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • Deubiquitinating enzyme 8 antibody
    • FLJ34456 antibody
    • hUBPy antibody
    • HumORF8 antibody
    • KIAA0055 antibody
    • MGC129718 antibody
    • SPG59 antibody
    • Ubiquitin carboxyl-terminal hydrolase 8 antibody
    • Ubiquitin isopeptidase Y antibody
    • ubiquitin specific peptidase 8 antibody
    • Ubiquitin thioesterase 8 antibody
    • Ubiquitin-specific-processing protease 8 antibody
    • UBP8_HUMAN antibody
    • UBPY antibody
    • Usp8 antibody
    see all

Images

  • Formalin-fixed, paraffin-embedded human breast carcinoma tissue stained for UBPY/USP8 using ab245641 at 1/200 dilution in immunohistochemical analysis. Detection: DAB staining.

  • All lanes : Anti-UBPY/USP8 antibody (ab245641) at 0.04 µg/ml

    Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 50 µg
    Lane 2 : HeLa whole cell lysate at 15 µg
    Lane 3 : HeLa whole cell lysate at 5 µg
    Lane 4 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 50 µg

    Developed using the ECL technique.

    Predicted band size: 128 kDa


    Exposure time: 3 minutes
  • UBPY/USP8 was immunoprecipitated from HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate (1 mg for IP, 20% of IP loaded) with ab245641 at 3 µg/mg lysate. Western blot was performed from the immunoprecipitate using ab245641 at 0.4 µg/ml.

    Lane 1: ab245641 IP in HeLa whole cell lysate.
    Lane 2: Control IgG IP in HeLa whole cell lysate.

    Detection: Chemiluminescence with exposure time of 10 seconds.

References

ab245641 has not yet been referenced specifically in any publications.

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