IL-18 ELISA Kit

Catalog No. ABIN1979353

Product Details IL-18 ELISA Kit

Target: IL-18 (IL18) (Interleukin 18 (IL18))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 0.5-75 pg/mL

Minimum Detection Limit: 0.5 pg/mL

Application: ELISA

Purpose: Human IL-18 ELISA Kit for cell culture supernatants, plasma, and serum samples.

Sample Type: Plasma, Cell Culture Supernatant, Serum

Analytical Method: Quantitative

Specificity: This ELISA kit shows no cross-reactivity with any of the cytokines tested: Human Angiogenin, BDNF, BLC, ENA-78, FGF-4, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, G-CSF, GM-CSF, IFN-gamma, Leptin, MCP-1, MCP-2, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, PARC, PDGF, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.

Cross-Reactivity (Details): This ELISA kit shows no cross-reactivity with any of the cytokines tested (e.g., human Angiogenin, BDNF, BLC, ENA-78, FGF-4, IL-1alpha, IL-1beta, IL-2, IL-3, IL-4, IL-5, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, IL-309, IP-10, G-CSF, GM-CSF, IFN-gamma, Leptin, MCP-1, MCP-2, MCP-3, MDC, MIP-1alpha, MIP-1 beta, MIP-1delta, PARC, PDGF, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF). X. TROUBLESHOOTING GUIDE Problem Cause Solution 1. Poor standard curve 1. Inaccurate pipetting 1. Check pipettes 2. Improper standard dilution 2. Ensure briefly spin the vial of Item C and dissolve the powder thoroughly by a gentle mix. 2. Low signal 1.Too brief incubation times 1. Ensure sufficient incubation time, assay procedure step 2 change to over night 2. Inadequate reagent volumes or improper dilution 2. Check pipettes and ensure correct preparation 3. Large CV 1. Inaccurate pipetting 1. Check pipettes 4. High background 1. Plate is insufficiently washed 1. Review the manual for proper wash. If using an a plate washer, check that all ports are unobstructed. 2. Contaminated wash buffer 2. Make fresh wash buffer 5. Low sensitivity 1. Improper storage of the ELISA kit 1. Store your standard at<-20oC after reconstitution, others at 4 oC. Keep substrate solution protected from light 2. Stop solution 2. Stop solution should be added to each well before measure The Human IL-18 ELISA Kit Protocol 10 The ELISA kits:5 Over 300 ELISA kits, custom ELISA kit choose from over 500 list visit www.raybiotech.com for details. The Human IL-18 ELISA Kit Protocol 11 The Human IL-18 ELISA Kit Protocol 12 This product is for research use only. ©2004 RayBiotech, Inc. The Human IL-18 ELISA Kit Protocol 13

Characteristics:

• Strip plates and additional reagents allow for use in multiple experiments
• Quantitative protein detection
• Establishes normal range
• The best products for confirmation of antibody array data

Components:

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent(s)
• Lyophilized Standard
• Biotinylated Detection Antibody
• Streptavidin-Conjugated HRP
• TMB One-Step Substrate

Material not included:

• Distilled or deionized water
• Precision pipettes to deliver 2 μL to 1 μL volumes
• Adjustable 1-25 μL pipettes for reagent preparation
• 100 μL and 1 liter graduated cylinders
• Tubes to prepare standard and sample dilutions
• Absorbent paper
• Microplate reader capable of measuring absorbance at 450nm
• Log-log graph paper or computer and software for ELISA data analysis

Application Details

Application Notes: Recommended Dilution for serum and plasma samples5 - 50 fold

Sample Volume: 100 μL

Plate: Pre-coated

Protocol:

1. Prepare all reagents, samples and standards as instructed in the manual.
2. Add 100 μL of standard or sample to each well.
3. Incubate 2.5 h at RT or O/N at 4 °C.
4. Add 100 μL of prepared biotin antibody to each well.
5. Incubate 1 h at RT.
6. Add 100 μL of prepared Streptavidin solution to each well.
7. Incubate 45 min at RT.
8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 μL of Stop Solution to each well.
11. Read at 450 nm immediately.

Reagent Preparation:

1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. 2. Sample dilution: If your samples need to be diluted, Assay Diluent A (Item D) should be used for dilution of serum/plasma samples, and 1x Assay Diluent B (Item E) should be used for dilution of culture supernatants and urine. Suggested dilution for normal serum/plasma: 5-50 fold. Please note that levels of

Assay Procedure:

1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. It is recommended that all standards and samples be run at least in duplicate. 2. Add 100 µL of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4 °C with gentle shaking. 3. Discard the solution and wash 4 times with 1x Wash Solution. Wash by filling each well with Wash Buffer (300 myl) using a multi-channel Pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels. 4. Add 100 µL of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking. 5. Discard the solution. Repeat the wash as in step 3. 6. Add 100 µL of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking. 7. Discard the solution. Repeat the wash as in step 3. 8. Add 100 µL of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking. The Human IL-18 ELISA Kit Protocol 6 9. Add 50 µL of Stop Solution (Item I) to each well. Read at 450 nm immediately.

Calculation of Results:

Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.

Assay Precision: Intra-Assay: CV<10%
Inter-Assay: CV<12%

Restrictions: For Research Use only

Handling

Handling Advice: Avoid repeated freeze-thaw cycles.

Storage: -20 °C

Storage Comment: The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.

Expiry Date: 6 months

References for IL-18 ELISA Kit (ABIN1979353)

Szpakowski, Biet, Locht, Paszkiewicz, Rudnicka, Druszczyńska, Allain, Fol, Pestel, Kowalewicz-Kulbat: "Dendritic Cell Activity Driven by Recombinant Mycobacterium bovis BCG Producing Human IL-18, in Healthy BCG Vaccinated Adults." in: Journal of immunology research, Vol. 2015, pp. 359153, (2016) (PubMed).

Kadoglou, Tahmatzidis, Giannakoulas, Kapelouzou, Gkontopoulos, Parissis, Lampropoulos, Kottas: "Serum levels of novel adipokines, omentin-1 and chemerin, in patients with acute myocardial infarction: KOZANI STUDY." in: Journal of cardiovascular medicine (Hagerstown, Md.), Vol. 16, Issue 5, pp. 341-6, (2015) (PubMed).

Al-Hakeim, Al-Rammahi, Al-Dujaili: "IL-6, IL-18, sIL-2R, and TNF? proinflammatory markers in depression and schizophrenia patients who are free of overt inflammation." in: Journal of affective disorders, Vol. 182, pp. 106-14, (2015) (PubMed).

Gileles-Hillel, Alonso-Álvarez, Kheirandish-Gozal, Peris, Cordero-Guevara, Terán-Santos, Martinez, Jurado-Luque, Corral-Peñafiel, Duran-Cantolla, Gozal: "Inflammatory markers and obstructive sleep apnea in obese children: the NANOS study." in: Mediators of inflammation, Vol. 2014, pp. 605280, (2014) (PubMed).

Mabrouk, Ghareeb, Shehab, Omar, El-Kabarity, Soliman, Mohamed: "Serum visfatin, resistin and IL-18 in A group of Egyptian obese diabetic and non diabetic individuals." in: The Egyptian journal of immunology / Egyptian Association of Immunologists, Vol. 20, Issue 1, pp. 1-11, (2013) (PubMed).

Jaiswal, Singh, Srivastava, Mittal: "Association of IL-12, IL-18 variants and serum IL-18 with bladder cancer susceptibility in North Indian population." in: Gene, Vol. 519, Issue 1, pp. 128-34, (2013) (PubMed).

Reichelt: "[The effect of ethionine on the enchondral ossification in rats]." in: Frankfurter Zeitschrift für Pathologie, Vol. 76, Issue 1, pp. 41-51 (PubMed).

Target Details for IL-18

Target: IL-18 (IL18) (Interleukin 18 (IL18))

Alternative Name: IL-18 (IL18 Products)

Synonyms: Igif ELISA Kit, Il-18 ELISA Kit, IL-18 ELISA Kit, IGIF ELISA Kit, ChIL-18 ELISA Kit, IL-1g ELISA Kit, IL1F4 ELISA Kit, IL18 ELISA Kit, interleukin 18 ELISA Kit, Il18 ELISA Kit, IL18 ELISA Kit

Background: IL-18 is one of a number of pro-inflammatory cytokines. The activities of IL-18 appear to be species specific. An important function of IL- 18 is the regulation of functionally distinct subsets of T-helper cells required for cell mediated immune responses. IL-18 functions as a growth and differentiation factor for Th1 cells. It is produced during the acute immune response by macrophages and immature dendritic cells. IL-18 is expressed by a variety of immune and non-immune cells.

Gene ID: 3606

UniProt: Q14116

Pathways: Cellular Response to Molecule of Bacterial Origin, Activated T Cell Proliferation, Cancer Immune Checkpoints, Inflammasome