|Product Name||Anti-IL18 Antibody|
|Storage & Handling||At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.|
|Description||Rabbit IgG polyclonal antibody for Interleukin-18(IL18) detection. Tested with WB, IHC-P, ELISA in Mouse.|
|Cite This Product||Anti-IL18 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # RP1017)|
|Immunogen||E. coli-derived mouse IL-18 recombinant protein(Position: N36-S192).|
Assay Dilutions Overview
ELISA , 0.1-0.5μg/ml, Mouse, -
Western blot, 0.1-0.5μg/ml, Mouse
Boster's Secondary Antibodies And IHC, WB Kits
The following reagents are used to generate the images below.Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P).
Images And Assay Conditions
Figure 1. Western blot analysis of IL-18 using anti- IL-18 antibody (RP1017).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: Recombinant Mouse IL18 Protein 10ng,
Lane 2: Recombinant Mouse IL18 Protein 5ng,
Lane 3: Recombinant Mouse IL18 Protein 2.5ng,
Lane 4: Recombinant Mouse IL18 Protein 1.25ng.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti- IL-18 antigen affinity purified polyclonal antibody (Catalog # RP1017) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for IL-18 at approximately 21KD. The expected band size for IL-18 is at 21KD.
Figure 2. IHC analysis of IL-18 using anti- IL-18 antibody (RP1017).
IL-18 was detected in paraffin-embedded section of mouse intestine tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti- IL-18 Antibody (RP1017) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Protein Target Info (Source: Uniprot.org)
|Alternative Names||Interleukin-18;IL-18;Interferon gamma-inducing factor;IFN-gamma-inducing factor;Interleukin-1 gamma;IL-1 gamma;Il18;Igif;|
|Molecular Weight||22135 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein Function||Augments natural killer cell activity in spleen cells and stimulates interferon gamma production in T-helper type I cells.|
|Background||Interleukin-18 also known as IL18 is a protein which in humans is encoded by the IL18 gene. The protein encoded by this gene is a proinflammatory cytokine.IL-18 is a cytokine produced by macrophages and other cells that belongs to the IL-1 superfamily. IL-18 works by binding to the interleukin-18 receptor, and together with IL-12 it induces cell-mediated immunity following infection with microbial products like lipopolysaccharide(LPS). After stimulation with IL-18, natural killer(NK) cells and certain T cells release another important cytokine called interferon-gamma(IFN-gamma) or type II interferon that plays an important role in activating the macrophages or other cells.The combination of this cytokine and IL12 has been shown to inhibit IL4 dependent IgE and IgG1 production, and enhance IgG2a production in B cells. IL-18 binding protein(IL18BP) can specifically interact with this cytokine, and thus negatively regulate its biological activity. The human interleukin 18 gene IL18 maps to 11q22.2-q22.3, closely linked to the DRD2 gene locus and distinct from mapped IDDM loci.|
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1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected
2. Post-translational cleavage: this can cause smaller bands and or multiple bands
3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody.
4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.
5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher.,