Overview

  • Product name
  • Description
    Rabbit polyclonal to HERC5
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Rabbit
  • Immunogen

    Synthetic peptide corresponding to Human HERC5 aa 900-949 (internal sequence).
    Sequence:

    FHPEELKDVIVGNTDYDWKTFEKNARYEPGYNSSHPTIVMFWKAFHKLTL

  • Positive control
    • HepG2 cell lysate.
  • General notes

    Protein previously labeled as HECT E3 ubiquitin ligase.

Properties

Applications

Our Abpromise guarantee covers the use of ab83853 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
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 117 kDa (predicted molecular weight: 117 kDa). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
IHC-P 1/100.

Target

  • Function
    Major E3 ligase for ISG15 conjugation. Acts as a positive regulator of innate antiviral response in cells induced by interferon. Makes part of the ISGylation machinery that recognizes target proteins in a broad and relatively non-specific manner. Catalyzes ISGylation of IRF3 which results in sustained activation, it attenuates IRF3-PIN1 interaction, which antagonizes IRF3 ubiquitination and degradation, and boosts the antiviral response. Catalyzes ISGylation of influenza A viral NS1 which attenuates virulence; ISGylated NS1 fails to form homodimers and thus to interact with its RNA targets. Catalyzes ISGylation of papillomavirus type 16 L1 protein which results in dominant-negative effect on virus infectivity. Physically associated with polyribosomes, broadly modifies newly synthesized proteins in a cotranslational manner. In an interferon-stimulated cell, newly translated viral proteins are primary targets of ISG15.
  • Tissue specificity
    Expressed in testis and to a lesser degree in brain, ovary and placenta. Found in most tissues at low levels.
  • Sequence similarities
    Contains 1 HECT (E6AP-type E3 ubiquitin-protein ligase) domain.
    Contains 5 RCC1 repeats.
  • Post-translational
    modifications
    ISGylated.
  • Cellular localization
    Cytoplasm > perinuclear region. Associated with the polyribosomes, probably via the 60S subunit.
  • Information by UniProt
  • Database links
  • Alternative names
    • CEB1 antibody
    • CEBP1 antibody
    • Cyclin E binding protein 1 antibody
    • Cyclin-E-binding protein 1 antibody
    • E3 ISG15--protein ligase HERC5 antibody
    • HECT domain and RCC1 like domain containing protein 5 antibody
    • HECT domain and RCC1-like domain-containing protein 5 antibody
    • Hect domain and RLD 5 antibody
    • HECT E3 ubiquitin ligase antibody
    • HERC5 antibody
    • HERC5_HUMAN antibody
    • Herc6 antibody
    see all

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human pineal tissue labelling HERC5 with ab83853 at 1/100. A Cy3-conjugated donkey anti-rabbit IgG (1/200) was used as the secondary antibody. Positive staining shown in the cytoplasm of cell bodies and processes of pinealocytes. Magnification: 20X. Exposure time: 0.5 - 2.0 seconds. Left - DAPI. Middle - HECT E3 ubiquitin ligase. Right - Merge.

  • Anti-HERC5 antibody (ab83853) at 1 µg/ml (5% skim milk / PBS ) + HepG2 cell lysate at 10 µg

    Secondary
    HRP conjugated anti-Rabbit IgG at 1/50000 dilution

    Predicted band size: 117 kDa
    Observed band size: 117 kDa
    Additional bands at: 55 kDa. We are unsure as to the identity of these extra bands.



    Gel concentration: 6-18%
  • ICC/IF image of ab83853 stained HeLa cells. The cells were 4% formaldehyde (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab83853, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References

ab83853 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Thank you for getting back to me with your progress.
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I think there are a few things that may be worthwhile trying in order to see if a better result may be obtained.
1. I would suggest using RIPA buffer to ma...

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Thank you for sharing that information. It has helped greatly in understanding the problems encountered in using the Anti-HECT E3 ubiquitin ligase antibody (ab83853).
I have ad a look at the protocols and I would have expected the antibody to prod...

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Thank you for contacting us. I am sorry to hear you are experiencing difficulties with the Anti-HECT E3 ubiquitin ligase antibody (ab83853). We take product complaints very seriously, and investigate every product that we feel may not be performing cor...

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