Human IL-17A (Interleukin 17 A) QuickTest ELISA Kit

This kit was based on sandwich ELISA method. The experiment lasted 120 minutes.

Capture antibody was conjugated to an affinity tag that was recognized by a specific antibody coated on the QuickTest plate.

Add the Cap/Det Ab working solution into each well, then add the standards and pilot samples into individual wells. If the sample contains IL-17, a capture antibody-IL-17-biotin-detection antibody complex was formed.

After incubation, unbound conjugates were removed by wash buffer. HRP-Streptavidin was added.

After washing, TMB substrates were added to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that turned yellow after adding a stop solution.

Read the O.D.

absorbance at 450nm in a microplate reader. The concentration of IL-17 in the sample was calculated by drawing a standard curve.

The concentration of the target substance is proportional to the OD450 value.

Catalogue No.: QT-EH3267

Alias: Interleukin-17A ELISA Kit, IL-17 ELISA Kit, IL-17A ELISA Kit, IL17A ELISA Kit

Species: Human

Range: 31.25-2000pg/ml

Sensitivity: 18.75pg/ml

Product NameHuman IL-17A (Interleukin 17 A) QuickTest ELISA KitAliasInterleukin-17A ELISA Kit, IL-17 ELISA Kit, IL-17A ELISA Kit, IL17A ELISA KitCatalogue No.QT-EH3267Size48T/96TSpeciesHumanUniProt No.

Q16552Sample TypeSerum, plasma, cell culture supernatant, cell lysate or tissue lysate, other biological fluid samplesDetection MethodSandwich ELISA, Double AntibodyDetection WavelengthOD450Reaction Duration120 minutesRange31.25-2000pg/mlSensitivity18.

75pg/mlStorage2-8°C(Sealed), Don't cryopreserve.SpecificitySpecifically binds with IL-17 , no obvious cross reaction with other analogues.

ELISA Kit Components

Kit Components — Item — Size(48T) — Size(96T) — Storage Condition for Opened Kit

E001 — ELISA
Microplate(Dismountable) — 8×6 — 8×12 — Put the rest strips into a sealed foil bag with the desiccant. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C

E002 — Lyophilized Standard — 1vial — 2vial — Put the rest standards into a desiccant bag. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C

E054 — Cap/Det Ab
(Ready to use, blue) — 3ml — 6ml — 2-8°C (Avoid Direct Light)

E053 — HRP-Streptavidin
(Ready to use, orange) — 5ml — 10ml

E024 — TMB Substrate — 5ml — 10ml

E039 — Sample Dilution Buffer — 20ml — 20ml — 2-8°C

E026 — Stop Solution — 5ml — 5ml

E038 — Wash Buffer(25X) — 15ml — 30ml

E006 — Plate Sealer — 3 pieces — 5 pieces

E007 — Product Description — 1 copy — 1 copy

Required Instruments and Reagents

1. Microplate reader (wavelength: 450nm)
2. 37°C incubator (CO2 incubator for cell culture is not recommenced.)
3. Automated plate washer or multi-channel pipette/5ml pipettor (for manual washing purpose)
4. Precision single (0.5-10μL, 5-50μL, 20-200μL, 200-1000μL) and multi-channel pipette with disposable tips(Calibration is required before use.)
5. Sterile tubes and Eppendorf tubes with disposable tips
6. Absorbent paper and loading slot
7. Deionized or distilled water

Assay Procedure Summary

• Step 1: Take out the required plate wells, add 50ul Cap/Det Ab into each well, then add 50ul 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 100ul 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 90ul 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 50ul stop solution. Read at 450nm immediately and calculate.

Standard CurveThis product is detected by QC department and meets performance required in the manual. (Laboratory Humidity: 20%-60%; Temperature: 18°C -25°C; Equilibrate TMB substrate to 37°C before staining.

After adding into the ELISA wells, incubate for 15min at 37°C in dark.)

Due to different assay environments and operations, assay data below and standard curve are provided for reference. Experimenters should establish standard curve according to their own assay.

STD.(pg/ml) — OD-1 — OD-2 — Average

0 — 0.071 — 0.075 — 0.073

31.25 — 0.13 — 0.131 — 0.13

62.5 — 0.175 — 0.175 — 0.175

125 — 0.281 — 0.284 — 0.269

250 — 0.476 — 0.482 — 0.453

500 — 0.694 — 0.714 — 0.716

1000 — 1.258 — 1.297 — 1.24

2000 — 2.175 — 2.168 — 2.144

RecoveryAdd a certain amount of IL-17 into the sample. Calculate the recovery by comparing the measured value with the expected amount of IL-17 in the sample.

Sample Type — Recovery Range(%) — Average(%)

serum(n=10) — 91-104 — 94

EDTA plasma(n=10) — 85-103 — 98

Heparin plasma(n=10) — 86-105 — 98

LinearityDilute the sample with a certain amount of IL-17 at 1:2, 1:4 and 1:8 to get the recovery range.

Sample Type — 1:2 — 1:4 — 1:8

serum(n=10) — 91-101% — 88-101% — 86-99%

EDTA plasma(n=10) — 82-100% — 85-98% — 85-100%

Heparin plasma(n=10) — 85-100% — 89-96% — 85-100%

Precision(%)Intra-assay Precision: samples with low, medium and high concentration are tested 20 times on the same plate.

Inter-assay Precision: samples with low, medium and high concentration are tested 20 times on three different plates.

Item — Intra-assay Precision — Inter-assay Precision

Sample — 1 — 2 — 3 — 1 — 2 — 3

n — 20 — 20 — 20 — 20 — 20 — 20

Mean (pg/ml) — 60.61 — 256.66 — 1005.41 — 63.24 — 253.41 — 971.38

Standard deviation — 2.67 — 10.11 — 38.91 — 2.73 — 10.54 — 44.78

CV(%) — 4.4 — 3.94 — 3.87 — 4.31 — 4.16 — 4.61

StabilityPerform the stability test for the sealed kit at 37°C and 2-8°C and get relevant data.

ELISA kit(n=5) — 37°C for 1 month — 2-8°C for 6 months

Average(%) — 80 — 95-100