Human IL-23(Interleukin 23) ELISA Kit

This kit is based on Double antibody-Sandwich ELISA detection method and takes 4h assay time. The microplate provided in this kit has been precoated with anti IL-23 antibody.

Add standard and properly diluted sample into relevant well respectively.

After incubation, wash unbound components. Add biotinylated detection antibody.

Then, it binds with IL-23 bound to precoated antibody. Wash unbound components and add HRP-Streptavidin Conjugate (SABC).

Wash unbound components again and add TMB substrate solution. Then, 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.

Calculate the concentration of IL-23 in the sample by plotting standard curve. The concentration of the target substance is proportional to the OD450 value.

Catalogue No.: EH3270

Alias: Interleukin-23 subunit alpha ELISA Kit, IL-23 subunit alpha ELISA Kit, IL-23-A ELISA Kit, Interleukin-23 subunit p19 ELISA Kit, IL-23p19 ELISA Kit, IL23A ELISA Kit, SGRF ELISA Kit

Species: Human

Range: 39.063-2500pg/ml

Sensitivity: 23.438pg/ml

Product NameHuman IL-23(Interleukin 23) ELISA KitAliasInterleukin-23 subunit alpha ELISA Kit, IL-23 subunit alpha ELISA Kit, IL-23-A ELISA Kit, Interleukin-23 subunit p19 ELISA Kit, IL-23p19 ELISA Kit, IL23A ELISA Kit, SGRF ELISA KitCatalogue No.EH3270Size48T/96TSpeciesHumanUniProt No.

Q9NPF7Sample TypeSerum, Plasma, Cell Culture Supernatant, cell or tissue lysate, Other liquid samplesDetection MethodSandwich ELISA, Double AntibodyDetection WavelengthOD450Reaction Duration4 hoursRange39.063-2500pg/mlSensitivity23.

438pg/mlStorage2-8°C(Sealed), Don't cryopreserve.SpecificitySpecifically binds with IL-23 , 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

E003 — Biotin-labeled Antibody(Concentrated, 100X) — 60ul — 120ul — 2-8°C (Avoid Direct Light)

E034 — HRP-Streptavidin Conjugate(SABC, 100X) — 60ul — 120ul

E024 — TMB Substrate — 5ml — 10ml

E039 — Sample Dilution Buffer — 10ml — 20ml — 2-8°C

E040 — Antibody Dilution Buffer — 5ml — 10ml

E049 — SABC Dilution Buffer — 5ml — 10ml

E026 — Stop Solution — 5ml — 10ml

E038 — Wash Buffer(Concentrated, 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: Add 100ul standard or sample into each well, seal the plate and statically incubate for 90 minutes at 37°C.
• Washing: Wash the plate twice without immersing.
• Step 2: Add 100ul biotin-antibody working solution, seal the plate and statically incubate for 60 minutes at 37°C.
• Washing: Wash the plate three times and immerse for 1min each time.
• Step 3: Add 100ul HRP-Streptavidin Conjugate (SABC) working solution, seal the plate and statically incubate for 30 minutes at 37°C.
• Washing: Wash the plate five times and immerse for 1min each time.
• Step 4: Add 90ul TMB substrate solution, seal the plate and statically incubate for 10-20 minutes at 37°C. (Accurate TMB visualization control is required.)
• Step 5: 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 — 0.16 — 0.164 — 0.162

39.062 — 0.216 — 0.222 — 0.219

78.125 — 0.353 — 0.363 — 0.358

156.25 — 0.505 — 0.519 — 0.512

312.5 — 0.716 — 0.736 — 0.726

625 — 0.967 — 0.995 — 0.981

1250 — 1.48 — 1.522 — 1.501

2500 — 2.275 — 2.341 — 2.308

RecoveryAdd a certain amount of IL-23 into the sample. Calculate the recovery by comparing the measured value with the expected amount of IL-23 in the sample.

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

serum(n=10) — 84-104 — 93

EDTA plasma(n=10) — 86-105 — 93

Heparin plasma(n=10) — 87-101 — 95

LinearityDilute the sample with a certain amount of IL-23 at 1:2, 1:4 and 1:8 to get the recovery range.

Sample Type — 1:2 — 1:4 — 1:8

serum(n=10) — 89-105% — 86-99% — 82-90%

EDTA plasma(n=10) — 87-96% — 84-100% — 87-104%

Heparin plasma(n=10) — 90-101% — 82-100% — 87-99%

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) — 77.11 — 306.7 — 1220 — 74 — 311.8 — 1228

Standard deviation — 3.88 — 13.31 — 53.56 — 3.18 — 13.47 — 53.05

CV(%) — 5.03 — 4.34 — 4.39 — 4.3 — 4.32 — 4.32

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