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ISG15 Antibody (703132RP488) in ICC/IF
Immunofluorescent analysis of ISG15 was performed using 70% confluent untreated and mouse IFN alpha (500 U/ml for 24 h) treated RAW 264.7 cells . The cells were fixed with 4% paraformaldehyde for 10 minutes, stained with Wheat Germ Agglutinin, Alexa Fluor™ 647 for 10 mins and permeabilized with 0.1% Triton X-100 for 15 minutes and blocked with 2% BSA for 1 hour at room temperature. The cells were stained with ISG15 Recombinant Rabbit Monoclonal Antibody (1H9L21), Alexa Fluor™ Plus 488 (Product # 703132RP488, 1:100) and Lamin B1 Recombinant... View More
Please note: We are reviewing Western blot images included in the antibody testing data in our catalog, including those provided by third parties. Unless expressly labeled or annotated as “raw-unedited”, Western blot images included in the antibody testing data in our catalog may have been edited, optimized or otherwise adjusted for presentation.
Product Details
703132RP488
Applications
Tested Dilution
Publications
Product Specifications
Species Reactivity
Mouse
Host/Isotype
Rabbit
/ IgG
Expression System
Expi293
Class
Recombinant Monoclonal
Type
Antibody
Clone
1H9L21
Immunogen
Protein corresponding to mouse ISG15 [aa2-aa155]
Conjugate
Alexa Fluor™ Plus 488
Alexa Fluor™ Plus 488
Alexa Fluor™ Plus 488
Excitation/Emission Max
493/518 nm
View spectra
Form
Liquid
Concentration
1.0 mg/mL
Amount
50 µg
Purification
Protein A
Storage buffer
proprietary buffer, pH 6.8
Contains
0.008% Bromonitrodioxane, 0.008% Methylisothiazolone
Storage conditions
4°C, store in dark, DO NOT FREEZE!
Shipping conditions
Wet ice
RRID
Product Specific Information
Alexa Fluor™ Plus recombinant antibodies are conjugated using new, proprietary dye chemistry so you can generate stunning data. Alexa Fluor™ Plus antibodies represent an advancement in fluorescent conjugate technology. Alexa Fluor™ Plus antibodies provide brighter signal compared to leading Alexa Fluor™ antibodies, providing you with better signal-to-noise for your critical experiments. These antibodies show better specificity and lot-to-lot consistency as these are recombinant antibodies, generated by cloning specific genes for the desired antibodies into an expression vector and expressed in vitro.
Using conjugate solutions: Centrifuge the protein conjugate solution briefly in a microcentrifuge before use; add only the supernatant to the experiment. This step will help eliminate any protein aggregates that may have formed during storage, thereby reducing nonspecific background staining.
Applications Tested: This antibody has been tested by western blot, immunocytochemistry and flow cytometric analysis of 3T3-L1 and RAW 264.7 cells treated with mouse IFN alpha. This may be used for immunocytochemistry at 10 µg/mL and for flow cytometry at less than or equal to 0.25 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
Excitation: 494 nm; Emission: 519 nm; Laser: Blue Laser
Filtration: 0.2 µm post-manufacturing filtered.
Target Information
Ubiquitin-like protein ISG15 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. It is secreted from monocytes in response to type I interferons and causes natural killer (NK)-cell proliferation and an augmentation of non-MCH (major histocompatibility complex)-restricted cytotoxicity. Synthesis is stimulated by IFN-alpha or IFN-beta or IFN-omega, but not IFN-gamma. ISG15 expression is also induced by overexpression of interferon regulatory factors that participate in transcriptional regulation of IFN genes, and by influenza B virus. ISG15 is secreted by cell lines of monocyte, T-lymphocyte, B-lymphocyte, human fibroblasts, and epithelial origins. Enhancement of NK cell proliferation, augmentation of non-major histocompatibility complex-restricted cytotoxicity, and induction of IFN-gamma from T cells identify ISG15 as a member of the cytokine cascade and suggest that it may be responsible for amplifying and directing some of the immunomodulatory effects of IFN-alpha or IFN-beta. ISG15 has has also been shown to function intracellularly as a ubiquitin homolog. Mutations affecting the gene can result in Immunodeficiency 38, with basal ganglia calcification (IMD38).
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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Bioinformatics
Protein Aliases: G1p2; interferon, alpha-inducible protein; interferon-induced 15 kDa protein; interferon-induced 15-KDa protein; interferon-induced 17 kDa protein; interferon-stimulated protein (15 kDa); interferon-stimulated protein 15; ubiquitin cross-reactive protein; unnamed protein product
Gene Aliases: G1p2; IGI15; IP17; Irfp; UCRP
UniProt ID: (Mouse) Q64339
Entrez Gene ID: (Mouse) 100038882
regulation of immune system process
integrin-mediated signaling pathway
response to virus
response to bacterium
modification-dependent protein catabolic process
positive regulation of bone mineralization
negative regulation of protein ubiquitination
ISG15-protein conjugation
positive regulation of protein oligomerization
regulation of interferon-gamma production
positive regulation of interferon-beta production
positive regulation of interferon-gamma production
positive regulation of interleukin-10 production
response to type I interferon
defense response to bacterium
negative regulation of viral genome replication
innate immune response
positive regulation of erythrocyte differentiation
positive regulation of multicellular organismal process
defense response to virus
negative regulation of type I interferon-mediated signaling pathway
protein localization to mitochondrion
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