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    products/primary-antibodies/isg15-antibody-epr3446-ab133346.pdf

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Cell Biology Proteolysis / Ubiquitin Proteasome / Ubiquitin Ub-like Proteins
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-ISG15 antibody [EPR3446] (ab133346)

  • Datasheet
  • SDS
Submit a review Submit a question References (3)

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Western blot - Anti-ISG15 antibody [EPR3446] (ab133346)
  • Western blot - Anti-ISG15 antibody [EPR3446] (ab133346)
  • Immunocytochemistry/ Immunofluorescence - Anti-ISG15 antibody [EPR3446] (ab133346)
  • Anti-ISG15 antibody [EPR3446] (ab133346)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3446] to ISG15
  • Suitable for: WB, ICC/IF
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Recombinant Human ISG15 protein (ab78929)

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Overview

  • Product name

    Anti-ISG15 antibody [EPR3446]
    See all ISG15 primary antibodies
  • Description

    Rabbit monoclonal [EPR3446] to ISG15
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IFmore details
    Unsuitable for: Flow Cyt,IHC-P or IP
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Daudi whole cell lysate (ab3951), IFN treated wild type HAP1 cell lysate, HeLa (untreated) and HeLa treated with 10 ng/ml IFN- a1 for 16 hours whole cell lysates ICC/IF: HeLa cells
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.2
    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EPR3446
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteasome / Ubiquitin
    • Ub-like Proteins
    • Epigenetics and Nuclear Signaling
    • Ubiquitin & Ubiquitin Like Modifiers
    • Ubiquitin like Modifiers

Associated products

  • Alternative Versions

    • Anti-ISG15 antibody [EPR3446] - BSA and Azide free (ab248489)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HeLa whole cell lysate (ab29545)
    • Daudi whole cell lysate (ab3951)
  • Recombinant Protein

    • Recombinant Human ISG15 protein (ab78929)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab133346 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 15 kDa (predicted molecular weight: 18 kDa).
ICC/IF
1/100 - 1/250.
Notes
WB
1/1000 - 1/10000. Detects a band of approximately 15 kDa (predicted molecular weight: 18 kDa).
ICC/IF
1/100 - 1/250.
Application notes
Is unsuitable for Flow Cyt,IHC-P or IP.

Target

  • Function

    Ubiquitin-like protein that is conjugated to intracellular target proteins after IFN-alpha or IFN-beta stimulation. Its enzymatic pathway is partially distinct from that of ubiquitin, differing in substrate specificity and interaction with ligating enzymes. ISG15 conjugation pathway uses a dedicated E1 enzyme, but seems to converge with the Ub conjugation pathway at the level of a specific E2 enzyme. Targets include STAT1, SERPINA3G/SPI2A, JAK1, MAPK3/ERK1, PLCG1, EIF2AK2/PKR, MX1/MxA, and RIG-1. Deconjugated by USP18/UBP43. Shows specific chemotactic activity towards neutrophils and activates them to induce release of eosinophil chemotactic factors. May serve as a trans-acting binding factor directing the association of ligated target proteins to intermediate filaments. May also be involved in autocrine, paracrine and endocrine mechanisms, as in cell-to-cell signaling, possibly partly by inducing IFN-gamma secretion by monocytes and macrophages. Seems to display antiviral activity during viral infections.
    In response to IFN-tau secreted by the conceptus, may ligate to and regulate proteins involved in the release of prostaglandin F2-alpha (PGF), and thus prevent lysis of the corpus luteum and maintain the pregnancy.
  • Tissue specificity

    Detected in lymphoid cells, striated and smooth muscle, several epithelia and neurons.
  • Sequence similarities

    Contains 2 ubiquitin-like domains.
  • Cellular localization

    Cytoplasm. Secreted. UCRP conjugates seem to be noncovalently associated with the intermediate filaments and distributed in a punctate pattern. Also secreted.
  • Target information above from: UniProt accession P05161 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 9636 Human
    • Omim: 147571 Human
    • SwissProt: P05161 Human
    • Unigene: 458485 Human
    • Alternative names

      • G1P2 antibody
      • hUCRP antibody
      • IFI 15 antibody
      • IFI15 antibody
      • Interferon alpha inducible protein antibody
      • Interferon induced protein 15 antibody
      • Interferon induced protein IFI-15K antibody
      • Interferon stimulated protein 15 kDa antibody
      • Interferon, alpha-inducible protein (clone IFI-15K) antibody
      • Interferon-induced 15 kDa protein antibody
      • Interferon-induced 17 kDa protein antibody
      • Interferon-induced 17-kDa/15-kDa protein antibody
      • IP17 antibody
      • ISG 15 antibody
      • ISG15 antibody
      • ISG15 ubiquitin like modifier antibody
      • ISG15_HUMAN antibody
      • Ubiquitin cross-reactive protein antibody
      • Ubiquitin-like protein ISG15 antibody
      • UCRP antibody
      see all

    Images

    • Western blot - Anti-ISG15 antibody [EPR3446] (ab133346)
      Western blot - Anti-ISG15 antibody [EPR3446] (ab133346)

      Lane 1: Wild type HAP1 whole cell lysate (20 µg)
      Lane 2: ISG15 IFN treated wild type HAP1 whole cell lysate (20 μg)
      Lane 3: ISG15 (KO) whole cell lysate (20 µg)
      Lane 4: ISG15 (KO) IFN alpha treated whole cell lysate (20 µg)
      Lane 5: Daudi whole cell lysate (20 µg)
      Lanes 1 - 5: Merged signal (red and green). Green - ab133346 observed at 18 kDa. Red - loading control, ab18058, observed at 130 kDa.

      ab133346 was shown to specifically react with ISG15 in IFN alpha1 induced wild-type HAP1cells. Loss of signal was observed when knockout cell line was used. Wild-type and HAP1 IFN treated knockout samples were subjected to SDS-PAGE. Ab133346 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1000 dilution and 1/30000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-ISG15 antibody [EPR3446] (ab133346)
      Western blot - Anti-ISG15 antibody [EPR3446] (ab133346)
      All lanes : Anti-ISG15 antibody [EPR3446] (ab133346) at 1/2000 dilution

      Lane 1 : Untreated HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
      Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with 10 ng/ml IFN- a1 for 16 hours whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 18 kDa
      Observed band size: 17 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking/Diluting buffer and concentration: 5% NFDM/TBST

    • Immunocytochemistry/ Immunofluorescence - Anti-ISG15 antibody [EPR3446] (ab133346)
      Immunocytochemistry/ Immunofluorescence - Anti-ISG15 antibody [EPR3446] (ab133346)
      ImmunImmunofluorescent analysis of ISG15 in HeLa cells stained with ab133346 at a 1/100 dilution.
    • Anti-ISG15 antibody [EPR3446] (ab133346)
      Anti-ISG15 antibody [EPR3446] (ab133346)

    Protocols

    • Western blot protocols
    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (3)

    Publishing research using ab133346? Please let us know so that we can cite the reference in this datasheet.

    ab133346 has been referenced in 3 publications.

    • Zhou H  et al. TMEM2 binds to CSNK2A3 to inhibit HBV infection via activation of the JAK/STAT pathway. Exp Cell Res 400:112517 (2021). PubMed: 33582094
    • Sultana T  et al. Multiple Pathways To Avoid Beta Interferon Sensitivity of HIV-1 by Mutations in Capsid. J Virol 93:N/A (2019). PubMed: 31511380
    • Li P  et al. Reciprocal regulation of miR-1205 and E2F1 modulates progression of laryngeal squamous cell carcinoma. Cell Death Dis 10:916 (2019). PubMed: 31801947

    Customer reviews and Q&As

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