AB248489

Anti-ISG15 antibody [EPR3446] - BSA and Azide free

RabMAb
Recombinant
KO Validated
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(1 Publication)

Rabbit Recombinant Monoclonal ISG15 antibody. Carrier free. Suitable for ICC/IF, WB and reacts with Human samples. Cited in 1 publication.

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

3 Images

Immunocytochemistry/ Immunofluorescence - Anti-ISG15 antibody [EPR3446] - BSA and Azide free (AB248489)

ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-ISG15 antibody [EPR3446] - BSA and Azide free (AB248489)

This data was developed using ab133346, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of ISG15 in HeLa cells stained with ab133346 at a 1/100 dilution.

Lab

Western blot - Anti-ISG15 antibody [EPR3446] - BSA and Azide free (AB248489)

This data was developed using ab133346, the same antibody clone in a different buffer formulation.

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.

All lanes:

Western blot - Anti-ISG15 antibody [EPR3446] (<a href='/en-us/products/primary-antibodies/isg15-antibody-epr3446-ab133346'>ab133346</a>)

Predicted band size: 18 kDa

Observed band size: 18 kDa

false

Lab

Western blot - Anti-ISG15 antibody [EPR3446] - BSA and Azide free (AB248489)

This data was developed using ab133346, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-ISG15 antibody [EPR3446] (<a href='/en-us/products/primary-antibodies/isg15-antibody-epr3446-ab133346'>ab133346</a>) at 1/2000 dilution

Lane 1:

Untreated HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) treated with 10 ng/ml IFN- a1 for 16 hours whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 18 kDa

Observed band size: 17 kDa

false

Exposure time: 3min

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR3446

Isotype

IgG

Carrier free

Yes

Reacts with

Human

Applications

ICC/IF, WB

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "FlowCyt" : {"fullname" : "Flow Cytometry", "shortname":"Flow Cyt"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "" } } }

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Product details

ab248489 is the carrier-free version of ab133346.

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

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification

Storage buffer

pH: 7.2 - 7.4 Constituents: PBS

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

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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.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

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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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Publications (1)

Recent publications for all applications. Explore the full list and refine your search

Oncology letters 24:457 PubMed36380881

2022

Suppressive effect of YY1-mediated RGS22 regulation on the proliferation, migration and invasion of pancreatic ductal adenocarcinoma.

Applications

Unspecified application

Species

Unspecified reactive species

Shou-Ji Cao,Wan-Li Ge,Ling-Dong Meng,Qun Chen,Yi Miao,Kui-Rong Jiang,Jing-Jing Zhang

View all publications

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Anti-ISG15 antibody [EPR3446] - BSA and Azide free

Immunocytochemistry/ Immunofluorescence - Anti-ISG15 antibody [EPR3446] - BSA and Azide free (AB248489)

Western blot - Anti-ISG15 antibody [EPR3446] - BSA and Azide free (AB248489)

Western blot - Anti-ISG15 antibody [EPR3446] - BSA and Azide free (AB248489)

Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Publications (1)

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