• Product name

  • Description

    Rabbit polyclonal to HERC5
  • Host species

  • 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).


  • Positive control

    • HepG2 whole cell lysate (ab7900).
  • General notes

    Protein previously labeled as HECT E3 ubiquitin ligase.



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.


  • 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

  • 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


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

    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%
  • 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.

  • 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.


ab83853 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A


Thank you for getting back to me with your progress.
I am sorry to hear that the results have not improved following my suggestions. I can certainly arrange for a refund to be paid to you. I have the order number xxxxx (Purchase order number xxxx) for you, ordered on the 24th of May 2012. I can arrange for a refund to be paid, which would go directly onto the account which paid for this order, or a credit note (which can be used against future orders with Abcam). Alternatively, I can arrange for a replacement antibody to be sent of your choice from our catalogue.
I look forward to hearing how you would like to proceed.

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Thank you for providing this extra information and sorry for the delay in my response.
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 make sure the cells are lysed sufficiently and the protein remains solubilise as best possible. A RIPA buffer recipe can be found form the following link:
2. I would also suggest using Laemmli buffer to before heating and load the samples onto the SDS-gel. This includes 10% 2-mercaptoethanol. Using a reducing agent can be worthwhile even when not looking at membrane proteins. This is because without reducing reagent the native structure may not be denatured completely, which can lead to aggregation as well as the protein not running as expected on the gel.
3. I would alter the diluent buffer from 1xTBS+1% Tween 20 to reduce the tween content to 0.1% as higher concentrations may remove the antibody from the membrane. I would also add 0.1% milk blocking in the buffer. I would do the same with the secondary antibody and only use TBS with 0.1% Tween20 when washing the membrane.
4. I do think it would also be worthwhile to perform a "no primary" control just to check if the secondary antibody is contributing to the non-specific band observed.
Hopefully these suggestions will lead to some improvement in the results observed. I would be very interested to hear how you get on.
If you have any further questions, please do not hesitate to contact us again.

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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 produce better results. In order to understand what may be contributing to the problems could I ask a few further questions?
1. Has a loading control been used (such as actin or GAPDH) in order to ascertain the quality of the samples loaded?
2. Has a "no primary" control been performed? If so, what was the results of this?
3. Was the protein transfer verified?
4. Were the samples boiled prior to loading to the gel? What reducing agent if any was used?
I look forward to receiving your reply.

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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 correctly.
In order to investigate what may be contributing to the problems encountered would you mind providing a little more information in relation to the protocol used and the results obtained? To this end, I have attached a questionnaire, this should only take 5-10 minutes to complete. Once I have receive the completed questionnaires, I will look at the protocols used and see if there are any suggestions we can make that may improve the results. If you could include any images of your results that would be very helpful.
This information will also allow us to investigate this case internally and initiate any additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.
I look forward to receiving your reply.

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