IL-17 ELISA Kit
Quick Overview for IL-17 ELISA Kit (ABIN7671681)
Target
See all IL-17 (IL17) ELISA KitsReactivity
Detection Method
Method Type
Detection Range
Application
Sample Type
Grade
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Minimum Detection Limit
- 31.25 pg/mL
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Purpose
- Human IL-17A (Interleukin 17 A) QuickTest ELISA Kit
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Analytical Method
- Quantitative
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Specificity
- Specifically binds with IL-17 , 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 ND-35pg/mL Healthy plasma (EDTA, Citrate , heparin) (n=18) undiluted ND-37pg/mL Human peripheral blood mononuclear cells (2 x 10^6 cells/mL) were cultured in RPMI 1640 supplemented with 10 %FBS, 100 units/mL penicillin, , and 100 μg/mL streptomycin sulfate for 24 hours. The cell culture supernatants were assayed for levels of human IL-17A. undiluted ND Human peripheral blood mononuclear cells (2 x 10^6 cells/mL) were cultured in RPMI 1640 supplemented with 10 %FBS, 100 units/mL penicillin, and 100 μg/mL streptomycin sulfate, followed by stimulated with 10 μg/mL PHA for 24 hours. The cell culture supernatants were assayed for levels of human IL-17A. undiluted 187pg/mL 2. KD sample validation (Detect IL-17-KD PBMC cells): Sample Type Dilution Ratio Content Wild Human peripheral blood mononuclear cells (2 x 10^6 cells/mL) were cultured in RPMI 1640 supplemented with 10 %FBS, 100 units/mL penicillin, and 100 μg/mL streptomycin sulfate, followed by stimulated with 10 μg/mL PHA for 24 hours. The cell culture supernatants were assayed for levels of human IL-17A. undiluted 169pg/mL Human peripheral blood mononuclear cells (+siRNA) were cultured in RPMI 1640 supplemented with 10 %FBS, 100 units/mL penicillin, and 100 μg/mL streptomycin sulfate, followed by stimulated with 10 μg/mL PHA for 24 hours. The cell culture supernatants were assayed for levels of human IL-17A. undiluted 29pg/mL 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. HEK293-derived Human IL-17A protein Gly24-Ala155B. E. coli-derived Human IL-17A protein Ile20-Ala155
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Sensitivity
- 18.75 pg/mL
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Components
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- ELISA Microplate(Dismountable)
- Lyophilized Standard
- Cap/Det Ab (Ready to use
- blue)
- TMB Substrate
- Sample Dilution Buffer
- Stop Solution
- Plate Sealer
- Product Description
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Material not included
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- 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
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Application Notes
- Optimal working dilution should be determined by the investigator.
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Comment
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Sandwich ELISA, Double Antibody
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Sample Volume
- 50 μL
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Assay Time
- 2 h
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Plate
- Pre-coated
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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.
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Assay Procedure
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- 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.
- Wash the plate twice without immersion.
- Add 100 µL HRP-Streptavidin (orange) into each well, seal the plate and statically incubate for 30 minutes at 37 °C.
- Wash the plate five times without immersion.
- 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.).
- Add 50 µL stop solution. Read at 450nm immediately and calculate.
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Restrictions
- For Research Use only
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Handling Advice
- Avoid direct light exposure to antibody solutions
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Storage
- 4 °C
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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
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Expiry Date
- 12 months
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- IL-17 (IL17) (Interleukin 17 (IL17))
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Alternative Name
- IL-17
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Background
- Interleukin-17A, IL-17, IL-17A, IL17A
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UniProt
- Q16552
Target See all IL-17 (IL17) ELISA Kits
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