| Applications: |
ELISA |
| Reactivity: |
Human |
| Note: |
FOR SCIENTIFIC EDUCATIONAL RESEARCH USE ONLY (RUO). MUST NOT BE USED IN DIAGNOSTIC OR OTHER MEDICAL APPLICATIONS. |
| Sensitivity: |
18.75pg/mL |
| Detection Limit: |
31.25~2000pg/mL |
| Short Description: |
This ISG15 Sandwich ELISA is an in-vitro enzyme-linked immunosorbent assay for the measurement of samples in human cell culture supernatant, serum and plasma (EDTA, citrate, heparin). |
| Storage Instruction: |
If unopened the kit may be stored at 2-8°C for up to 1 month. If the kit will not be used within 1 month, store the components separately, according to the component table in the manual. |
| Assay Time: |
3.5h |
| Detection: |
Colormetric |
| Gene Symbol: |
ISG15 |
| Gene ID: |
9636 |
| Uniprot ID: |
ISG15_HUMAN |
| Specificity: |
This kit recognizes Human ISG15 in samples. No significant cross-reactivity or interference between Human ISG15 and analogues was observed. |
| Sample Type: |
Serum, plasma and other biological fluids |
| Post Translational Modifications | S-nitrosylation decreases its dimerization, thereby increasing the availability as well as the solubility of monomeric ISG15 for its conjugation to cellular proteins. Induced as an inactive, precursor protein that is cleaved by specific proteases to expose the C-terminal diglycine (LRLRGG) motif. This motif is essential not only for its conjugation to substrates but also for its recognition by the relevant processing proteases. |
| Function | Ubiquitin-like protein which plays a key role in the innate immune response to viral infection either via its conjugation to a target protein (ISGylation) or via its action as a free or unconjugated protein. ISGylation involves a cascade of enzymatic reactions involving E1, E2, and E3 enzymes which catalyze the conjugation of ISG15 to a lysine residue in the target protein. Its target proteins include IFIT1, MX1/MxA, PPM1B, UBE2L6, UBA7, CHMP5, CHMP2A, CHMP4B and CHMP6. Isgylation of the viral sensor IFIH1/MDA5 promotes IFIH1/MDA5 oligomerization and triggers activation of innate immunity against a range of viruses, including coronaviruses, flaviviruses and picornaviruses. Can also isgylate: EIF2AK2/PKR which results in its activation, RIGI which inhibits its function in antiviral signaling response, EIF4E2 which enhances its cap structure-binding activity and translation-inhibition activity, UBE2N and UBE2E1 which negatively regulates their activity, IRF3 which inhibits its ubiquitination and degradation and FLNB which prevents its ability to interact with the upstream activators of the JNK cascade thereby inhibiting IFNA-induced JNK signaling. Exhibits antiviral activity towards both DNA and RNA viruses, including influenza A, HIV-1 and Ebola virus. Restricts HIV-1 and ebola virus via disruption of viral budding. Inhibits the ubiquitination of HIV-1 Gag and host TSG101 and disrupts their interaction, thereby preventing assembly and release of virions from infected cells. Inhibits Ebola virus budding mediated by the VP40 protein by disrupting ubiquitin ligase activity of NEDD4 and its ability to ubiquitinate VP40. ISGylates influenza A virus NS1 protein which causes a loss of function of the protein and the inhibition of virus replication. The secreted form of ISG15 can: induce natural killer cell proliferation, act as a chemotactic factor for neutrophils and act as a IFN-gamma-inducing cytokine playing an essential role in antimycobacterial immunity. The secreted form acts through the integrin ITGAL/ITGB2 receptor to initiate activation of SRC family tyrosine kinases including LYN, HCK and FGR which leads to secretion of IFNG and IL10.the interaction is mediated by ITGAL. |
| Protein Name | Ubiquitin-Like Protein Isg15Interferon-Induced 15 Kda ProteinInterferon-Induced 17 Kda ProteinIp17Ubiquitin Cross-Reactive ProteinHucrp |
| Database Links | Reactome: R-HSA-1169408Reactome: R-HSA-168276Reactome: R-HSA-168928Reactome: R-HSA-5656169Reactome: R-HSA-909733Reactome: R-HSA-936440Reactome: R-HSA-9705671 |
| Cellular Localisation | CytoplasmSecretedExists In Three Distinct States: Free Within The CellReleased Into The Extracellular SpaceOr Conjugated To Target Proteins |
| Alternative ELISA Names | Ubiquitin-Like Protein Isg15 ELISA kitInterferon-Induced 15 Kda Protein ELISA kitInterferon-Induced 17 Kda Protein ELISA kitIp17 ELISA kitUbiquitin Cross-Reactive Protein ELISA kitHucrp ELISA kitISG15 ELISA kitG1P2 ELISA kitUCRP ELISA kit |
| Specificity | This kit recognizes Human ISG15 in samples. No significant cross-reactivity or interference between Human ISG15 and analogues was observed. |
| Reproducibility | Both intra-CV and inter-CV are |
Information sourced from Uniprot.org
| Item | Specifications | Storage |
| Micro ELISA Plate (Dismountable) | 96T: 8 wells ×12 strips strips | -20℃, 6 months |
| Reference Standard | 96T: 2 vials 48T: 1 vial | -20℃, 6 months |
| Concentrated Biotinylated Detection Ab (100×) | 96T: 1 vial, 120 μL 60 μL | -20℃, 6 months |
| Concentrated HRP Conjugate (100×) | 96T: 1 vial, 120 μL 60 μL | -20℃ (Protect from light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 2-8°C, 6 months |
| Biotinylated Detection Ab Diluent | 1 vial, 14 mL | 2-8°C, 6 months |
| HRP Conjugate Diluent | 1 vial, 14 mL | 2-8°C, 6 months |
| Concentrated Wash Buffer (25×) | 1 vial, 30 mL | 2-8°C, 6 months |
| Substrate Reagent | 1 vial, 10 mL | 2-8℃ (Protect from light) |
| Stop Solution | 1 vial, 10 mL | 2-8°C |
| Plate Sealer | 5 pieces | |
| Manual | 1 copy | |
| Certificate of Analysis | 1 copy | |
| Sample Type | Range (%) | Average Recovery (%) |
| Serum(n=8) | 89-102 | 95 |
| EDTA plasma (n=8) | 94-107 | 101 |
| Cell culture media (n=8) | 84-99 | 91 |
| | Intra-assay Precision | Intra-assay Precision | Intra-assay Precision | Inter-assay Precision | Inter-assay Precision | Inter-assay Precision |
| Sample | 1.00 | 2.00 | 3.00 | 1.00 | 2.00 | 3.00 |
| n | 20.00 | 20.00 | 20.00 | 20.00 | 20.00 | 20.00 |
| Mean (pg/mL) | 99.10 | 288.84 | 966.26 | 89.56 | 303.51 | 1026.46 |
| Standard deviation | 6.05 | 11.81 | 51.41 | 5.90 | 12.44 | 40.44 |
| CV (%) | 6.10 | 4.09 | 5.32 | 6.59 | 4.10 | 3.94 |
12 months for antibodies. 6 months for ELISA Kits. Please see website T&Cs for further guidance
