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AB303764

Human ISG15 ELISA Kit

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Human ISG15 ELISA Kit is a Sandwich (quantitative) ELISA for the measurement of Human ISG15 in Human in Cell/Tissue Extracts samples.

View Alternative Names

G1P2, UCRP, ISG15, Ubiquitin-like protein ISG15, Interferon-induced 15 kDa protein, Interferon-induced 17 kDa protein, Ubiquitin cross-reactive protein, IP17, hUCRP

2 Images
Sandwich ELISA - Human ISG15 ELISA Kit (AB303764)
  • sELISA

Supplier Data

Sandwich ELISA - Human ISG15 ELISA Kit (AB303764)

Interpolated concentration of native ISG15 was measured in duplicate at different sample concentrations. Undiluted samples are 5 µg/mL Hela treated and HT-29 cell extracts. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Sample dilutions are made in 1X Cell Extraction Buffer PTR

Sandwich ELISA - Human ISG15 ELISA Kit (AB303764)
  • sELISA

Supplier Data

Sandwich ELISA - Human ISG15 ELISA Kit (AB303764)

Example of human ISG15 standard curve. Background-subtracted data values (mean +/- SD) are graphed.

Key facts

Detection method

Colorimetric

Sample types

Cell Lysate

Reacts with

Human

Assay type

Sandwich (quantitative)

Sensitivity

= 3.201 pg/mL

Range

28.125 - 1800 pg/mL

Assay time

1h 30m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "sELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

Product details

Human ISG15 SimpleStep ELISA® kit (ab303764) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Human ISG15 protein in cell extracts and tissue extracts. Quantitate Human ISG15 with 3.201 pg/ml sensitivity.

SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:

-Single-wash protocol reduces assay time to 90 minutes or less
-High sensitivity, specificity and reproducibility from superior antibodies
-Fully validated in biological samples
-96-wells plate breakable into 12 x 8 wells strips

A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Precision

[ { "reproducibilityType": "Intra", "sample": "Extract", "replicates": 8, "mean": null, "standardDeviation": null, "coefficientOfVariability": "3.1" }, { "reproducibilityType": "Inter", "sample": "Extract", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "1.2" } ]

Recovery

[ { "sample": "Cell Lysate", "range": "89 - 94 %", "average": "= 91" } ]

What's included?

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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

ISG15 also known as G1P2 is a ubiquitin-like protein with a molecular weight of approximately 17 kDa. This protein gets expressed highly in response to interferon stimulation particularly in white blood cells such as macrophages and monocytes. It consists of two ubiquitin-like domains contributing to its functionality. ISG15 performs its mechanical role by adding itself to target proteins in a process known as ISGylation. This process resembles ubiquitination and serves to modify the functions of targeted proteins.
Biological function summary

ISG15 plays an important role in the immune response to viral infections. It acts as an antiviral agent by inhibiting viral replication. It interacts with and becomes a part of protein complexes modifying cellular proteins involved in immune regulation. ISG15 influences the stability function and localization of these target proteins enhancing antiviral defenses and potentially modulating immune signaling pathways.

Pathways

ISG15 operates within the interferon signaling pathway and the antiviral response pathway. In the interferon signaling pathway ISG15 interacts with proteins like UBE1L and UBE2L6 which act together for effective ISGylation of targets. In the antiviral response pathway ISG15 boosts the effects of proteins like IRF3 and STAT1 enhancing antiviral responses. These interactions allow ISG15 to effectively contribute to halting viral propagation.

ISG15 connects to autoimmune diseases and certain viral infections. In autoimmune conditions like systemic lupus erythematosus abnormal ISG15 activity can contribute to disease pathology. ISG15 also interacts with proteins such as USP18 which regulates the expression of type I interferons to influence the disease processes. Furthermore ISG15 involvement in viral infections such as hepatitis C highlights its value in antiviral defenses. Understanding these interactions can support the development of therapies targeting ISG15 for managing immune-related conditions.

Product protocols

Target data

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 (PubMed : 27564865). 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 (PubMed : 33727702). 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 (PubMed : 33727702). 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 (PubMed : 29100055).
See full target information ISG15
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