Product nameAnti-ISG15 antibody [EPR3446]
See all ISG15 primary antibodies
DescriptionRabbit monoclonal [EPR3446] to ISG15
Tested applicationsSuitable for: WB, ICC/IFmore details
Unsuitable for: Flow Cyt,IHC-P or IP
Species reactivityReacts with: Human
Synthetic peptide corresponding to residues of Human ISG15 (UniProt: P05161).
- 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
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.
This product is a recombinant rabbit monoclonal antibody.
Storage instructionsShipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
Storage bufferpH: 7.40
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol, 9.85% Tris glycine, 50% Tissue culture supernatant
PurityTissue culture supernatant
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.
|WB||1/1000 - 1/10000. Detects a band of approximately 15 kDa (predicted molecular weight: 18 kDa).|
|ICC/IF||1/100 - 1/250.|
FunctionUbiquitin-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 specificityDetected in lymphoid cells, striated and smooth muscle, several epithelia and neurons.
Sequence similaritiesContains 2 ubiquitin-like domains.
Cellular localizationCytoplasm. Secreted. UCRP conjugates seem to be noncovalently associated with the intermediate filaments and distributed in a punctate pattern. Also secreted.
- Information by UniProt
- G1P2 antibody
- hUCRP antibody
- IFI 15 antibody
Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: HAP1 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 HAP1 IFN treated when HAP1 IFN treated knockout samples were 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.
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.
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
ImmunImmunofluorescent analysis of ISG15 in HeLa cells stained with ab133346 at a 1/100 dilution.
ab133346 has not yet been referenced specifically in any publications.