IL-18 ELISA Kit

Catalog No. ABIN7671688

The Human IL-18 ELISA Kit (ABIN7671688) is a Colorimetric ELISA Kit designed to quantify Human IL-18.

Quick Overview for IL-18 ELISA Kit (ABIN7671688)

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

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 15.625 pg/mL - 1000 pg/mL

Application: ELISA

Sample Type: Biological Fluids, Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Lysate

Grade: Quick

Product Details IL-18 ELISA Kit

Minimum Detection Limit: 15.625 pg/mL

Purpose: Human IL-18 (Interleukin 18) QuickTest ELISA Kit

Analytical Method: Quantitative

Specificity: Specifically binds with IL-18 , no obvious cross reaction with other analogues. Recommended Sample Dilution Ratio The following table shows the recommended dilution ratios for this kit for a limited number of samples for your reference only. (When the sample content is very low, the original solution can be added without dilution, but it is necessary to ensure that the pH is between 6.8 and 8.0, and it does not contain more than 10 % organic solvents or high-concentration protein denaturants.) 1. Common sample validation: Sample Type Recommended Dilution Ratio Content Healthy serum (n=20) undiluted 17-355pg/mL Healthy plasma (EDTA, Citrate , heparin) (n=22) undiluted 18-386pg/mL THP-1 cells were cultured in RPMI 1640 supplemented with 80 nM TPA, 10 %FBS, 100 units/mL penicillin, and 100 μg/mL streptomycin sulfate, incubated overnight. The cell culture supernatants were assayed for levels of human IL-18. undiluted ND THP-1 cells were cultured in RPMI 1640 supplemented with 80 nM TPA, 10 %FBS, 100 units/mL penicillin, and 100 μg/mL streptomycin sulfate, incubated overnight. Cells were then stimulated with 100 ng/mL LPS for 12 hours, and the cell culture supernatants were assayed for levels of human IL-18. undiluted 496pg/mL Hela cells were cultured in DMEM supplemented with 10 %FBS, 100 units/mL penicillin, and 100 μg/mL streptomycin sulfate for 12 hours. The cell culture supernatants were assayed for levels of human IL-18. undiluted 267pg/mL 2. KO sample validation (Detect IL-18 KO Hela cells): Sample Type Dilution Ratio Content Wild Hela cells were cultured in DMEM supplemented with 10 %FBS, 100 units/mL penicillin, and 100 μg/mL streptomycin sulfate for 12 hours. The cell culture supernatants were assayed for levels of human IL-18. undiluted 318pg/mL KO Hela cells were cultured in DMEM supplemented with 10 %FBS, 100 units/mL penicillin, and 100 μg/mL streptomycin sulfate for 12 hours. The cell culture supernatants were assayed for levels of human IL-18. undiluted ND Note:ND is lower than the sensitivity of the kit and was not detected 3. Verified recombinant protein: (It is a normal phenomenon that some proteins have weak detection signals or cannot be detected at all due to differences in tags, sequences or protein activities) A.E. coli-derived Human IL-18 protein Tyr37-Asp193

Sensitivity: 9.375 pg/mL

Components:

• ELISA Microplate(Dismountable)
• Lyophilized Standard
• Cap/Det Ab (Ready to use
• blue)
• TMB Substrate
• Sample Dilution Buffer
• Stop Solution
• Plate Sealer
• Product Description

Material not included:

• Microplate reader (wavelength: 450nm)
• 37 °C incubator (CO2 incubator for cell culture is not recommenced.)
• Automated plate washer or multi-channel pipette/5 mL pipettor
• Precision single (0.5-10μL
• 5-50μL
• 20-200μL
• 200-1000μL) and multi-channel pipette with disposable tips(Calibration is required before use.)
• Sterile tubes and Eppendorf tubes with disposable tips
• Absorbent paper and loading slot
• Deionized or distilled water

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Comment:

Sandwich ELISA, Double Antibody

Sample Volume: 50 μL

Assay Time: 2 h

Plate: Pre-coated

Protocol: Step 1: Take out the required plate wells, add 50 µL Cap/Det Ab into each well, then add 50 µL Standard or Sample into individual well. (When adding standard or sample, the disposable tip lightly touches the liquid level. Change the disposable tips for different samples and standards.) Gently tap the plate for 10s to ensure thorough mixing then static incubate for 60 minutes at 37 °C. Washing: Wash the plate twice without immersion. Step 2: Add 100 µL HRP-Streptavidin (orange) into each well, seal the plate and static incubate for 30 minutes at 37 °C. Washing: Wash the plate five times without immersion. Step 3: Add 90 µL TMB substrate solution, seal the plate and static incubate for 10-20 minutes at 37 °C. (Accurate TMB visualization control is required.) Step 4: Add 50 µL stop solution. Read at 450nm immediately and calculate.

Assay Procedure:

1. Take out the required strips, add 50 µL Cap/Det Ab into each well, then add 50 µL Standard or Sample into relevant well. (When adding standard or sample, the disposable tip lightly touches the liquid level. Change the disposable tips for different samples and standards.) Gently tap the plate for 10s to ensure complete mixing then statically incubate for 60 minutes at 37 °C.
2. Wash the plate twice without immersion.
3. Add 100 µL HRP-Streptavidin (orange) into each well, seal the plate and statically incubate for 30 minutes at 37 °C.
4. Wash the plate five times without immersion.
5. Add 90 µL TMB substrate solution, seal the plate and statically incubate for 10-20 minutes at 37 °C (Accurate TMB visualization control is required.).
6. Add 50 µL stop solution. Read at 450nm immediately and calculate.

Restrictions: For Research Use only

Handling

Handling Advice: Avoid direct light exposure to antibody solutions

Storage: 4 °C

Storage Comment: Store sealed Kit at 2-8° C. Do Not Cryopreserve. Store ELISA Microplate for 1 month at 2-8°C, Store for 12 month at -20°C. Put the leftover strips into a sealed foil bag with the desiccant

Expiry Date: 12 months

Target Details for IL-18

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

Alternative Name: IL-18

Background: Interleukin-18, IL-18, Iboctadekin, Interferon gamma-inducing factor, IFN-gamma-inducing factor, Interleukin-1 gamma, IL-1 gamma, IL18, IGIF, IL1F4

UniProt: Q14116

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