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Antibodies

IL-18 Antibody

Product Sizes
25 ul
210-401-323S-25UL
About this Product
SKU:
210-401-323S
Additional Names:
Il18|rabbit anti-IL-18 antibody, rabbit anti-interleukin-18 antibody, Iboctadekin antibody, IFN gamma inducing factor antibody, IGIF antibody, IL 1 gamma antibody, IL 18 antibody, IL 1g antibody, IL-1F4, IL 18, Interleukin 18, IL18, Interleukin18, IL1 F4, IL1F4, Ms IL-18, mouse IL18
Application:
ELISA, Immunofluorescence, Immunohistochemistry, Western Blot
Buffer:
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride
CE/IVD:
RUO
Clonality:
Polyclonal
Concentration:
1 mg/ml
Extra Details:
Interleukin-18 (IL-18) is a member of the IL-1 cytokine family and was initially identified as an Interferon-g (IFN-g) inducing factor (IGIF). The IL-18 gene was originally cloned from liver cells and has since been shown to be produced by activated monocytes/ macrophages, Kupffer cells, keratinocytes, glucocorticoid-secreting adrenal cortex cells, osteoblasts and dendritic cells. IL-18 is a 24 kDa, non-glycosylated polypeptide that lacks a classical signal sequence and possesses a structure recognizably similar to IL-1. IL-18 is synthesized as a bio-inactive propeptide that undergoes proteolytic cleavage by either ICE (interleukin-1 beta converting enzyme) or another caspase to generate a mature, bioactive, 18 kDa molecule. In both the mature and propeptide forms, IL-18 shows 64% aa sequence identity from mouse to human. IL-18 does not appear to show any primary sequence similarity to any other known cytokines. Rat IL-18 has also been isolated, and found to be 194 aa in length with a 91% aa sequence identity to mouse IL-18. Human IL-18 has been found to induce the production of IFN-g and GM-CSF while inhibiting the production of IL-10 by PBMC. With respect to human T cells, IL-18 enhances Th1 cytokine production and stimulates cell proliferation via an IL-2-dependent pathway. Human IL-18 can also inhibit the synthesis of IgE by B cells. Thus, IL-18 plays an important role in immunological and inflammatory reactions. Currently, the bioactivity of human IL-18 is often determined by its capacity to augment the levels of IFN-g produced by T cells as measured in tissue culture supernatants.
Formulation:
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Host:
Rabbit
Immunogen:
The whole rabbit serum used to produce this IgG fraction antibody was prepared by repeated immunizations with native 157 aa mouse IL-18 produced in E.coli.
Isotype:
IgG
Physical State:
Liquid
Reactivities:
Mouse
Shipping Conditions:
Dry Ice
Specificity:
This is an IgG preparation of whole rabbit serum purified by protein A chromatography using a low endotoxin methodology. This antibody is primarily directed against mature 18,000 MW mouse IL-18 and is useful in determining its presence in various assays. This antibody will also recognize the 24,000 inactive precursor form of mouse IL-18. In general, this antibody also detects rat IL-18 in the same
Storage Conditions:
-20[o]C diluted. Avoid freeze/thaw cycles., See Manual Avoid freeze/thaw cycles.
Supplier:
Rockland Inc
Type:
Antibody: Polyclonal Antibody
Immunohistochemistry with Rabbit anti-Mouse IL-18 antibody showing IL-18 staining in inflammatory cells of the mucous corium of mouse colon at 20x and 40x. Slide A is a negative control. Slides B and C show staining. Formalin fixed/paraffin embedded sections were subjected to heat induced epitope retrieval (HIER) at pH 6.2 and then incubated with mouse anti-IL-18 antibody at 4.0 ug/ml for 60 minutes. The reaction was developed using MACH 4 universal AP polymer detection system and visualized with WARP RED.
In-vitro neutralization. Spleens were aseptically removed and cell suspensions were prepared. Cells were washed twice and resuspended in RPMI supplemented with 10% FBS. For cytokine measurement, spleen cells were cultured at 5 mln/mL in 24-well, flat-bottom culture plates in the presence of several dilutions of rabbit anti-murine IL-18 antibody (1:400; 1:200; 1:100; 1:50) and 100 ng/mL of LPS (a phenol-extracted preparation from Escherichia coli 055:B5, Sigma Chemical Co). Cultures were incubated at 37°C in a humidified atmosphere with 5% CO2. At the end of the incubation period, cultures were frozen at -70°C and subjected to 3 freeze-thaw cycles to obtain total cytokine levels. Before assaying, samples were centrifuged for 10 minutes at 10,000g to remove debris.
Immunofluorescence microscopy of IL-18 in mouse colon sections. The transversing portion of the large intestine from DSS-exposed (Panel A) and -unexposed mice (Panel B) was excised, rinsed in PBS, and frozen on isopentane cooled with liquid nitrogen. Frozen sections (5 um) were cut on a Leica CM 1850 cryostat. The slides were fixed for 10 min in 4% paraformaldehyde, air-dried, and incubated for 20 min in PBS supplemented with 10% normal goat serum. Sections were incubated in a 1:50 dilution of Rockland's rabbit anti-Mouse IL-18 antibody or 1 ug/ml nonimmune rabbit IgG (not shown) as negative control. The antibodies were diluted in PBS containing 1% bovine serum albumin. After an overnight incubation at 4°C, the sections were washed three times with 0.5% bovine serum albumin in PBS. The sections were then incubated with a secondary goat anti-rabbit antibody conjugated to Alexa488 (Molecular Probes) for 60 min at room temperature in the dark. Nuclei were counterstained blue using 1 ug/100 ml bisbenzimide. After staining, sections were washed and examined with the Leica DM RXA confocal laser scanning system and analyzed. Similar staining will occur with other systems.