Technical Manual for Human Interleukin-23 (IL-23) ELISA Detection Kit

BS-0194 human interleukin-23 (IL-23) ELISA kit, native assay kit

1、 Product Overview

This kit uses double antibody sandwich enzyme-linked immunosorbent assay (ELISA) technology, specifically designed for quantitative detection of interleukin-23 (IL-23) concentration in human serum, plasma, tissue homogenate, and cell supernatant. The core components include enzyme-linked immunosorbent assay (ELISA) plates pre coated with antibodies, standards, biotinylated detection of antibodies, and chromogenic substrates (TMB), ensuring high sensitivity (as low as 0.15 pg/mL) and specificity. The entire experiment lasts about 3.5 hours and is suitable for scientific research experiments. Clinical diagnostic purposes are strictly prohibited. ‌

2、 Detection principle

Antibody binding: IL-23 binds to pre packaged captured antibodies to form a complex. ‌

Add biotinylated detection antibodies and horseradish peroxidase labeled streptavidin, and develop color through enzymatic reaction.

Quantitative analysis: After TMB substrate color development, the reaction was terminated by terminating the solution. The absorbance value was measured at 450 nm wavelength, and the concentration of IL-23 was positively correlated with the OD value. ‌

3、 Composition of reagent kit

|Components | Specifications/Quantity | Storage Conditions|

|-||-|

|Pre packaged enzyme-linked immunosorbent assay plate | 96 wells/plate | 2-8 ℃, away from light|

|Standard product (freeze-dried powder) | 2 bottles | Long term storage at -20 ℃|

|Biotin labeled antibody | 1 tube (concentrated solution) | 2-8 ℃ short-term|

|Chromogenic substrate (TMB) | 12 mL | 2-8 ℃, avoid light|

|Termination solution | 12 mL | Room temperature|

|20 x detergent | 50 mL | room temperature|

Note: The sample diluent should be selected according to the sample type (serum/plasma/cell supernatant).

4、 Sample processing

Serum/plasma: coagulate at room temperature for 30 minutes, centrifuge at 1000 × g for 15 minutes, and take the supernatant to avoid hemolysis.

Cell supernatant: Remove cell debris directly by centrifugation. Untested samples should be packaged and stored at -20 ℃ to avoid repeated freezing and thawing.

Tissue homogenate: Add lysis buffer homogenate according to the weight of the tissue and centrifuge to obtain the supernatant. ‌

5、 Operation steps

Reagent equilibrium: All components are allowed to stand at room temperature for 20 minutes, and concentrated washing solution is used to dissolve the crystals in a water bath. ‌

Sample addition reaction:

Add the standard sample to the enzyme labeled well after gradient dilution, with a sample volume of 25 μ L. ‌

Add the detection antibody and enzyme labeled complex in sequence, and incubate at 37 ℃ for 60 minutes.

Washing color development: Wash with washing solution three times, add TMB substrate and react in the dark for 15 minutes, then stop with the termination solution. ‌

Reading: The OD value is measured at a wavelength of 450 nm using an enzyme-linked immunosorbent assay (ELISA) reader, and the concentration is calculated by fitting four parameters.

6、 Performance parameters

Detection range: 15.6-1000 pg/mL (linear interval).

Sensitivity: 0.15 pg/mL.

Precision: The coefficient of variation (CV) within/between boards is less than 10%. ‌

Recovery rate: 80% -120%.

7、 Precautions

Safety protection: Wear gloves and goggles during operation to avoid contact with termination solution and TMB substrate. ‌

Reagent storage: unopened reagent kit can be stored for 6 months at 2-8 ℃, and for 1 year at -20 ℃; The opened reagent needs to be used up within 7 days.

Experimental specifications:

Mixing different batches of reagents is prohibited. ‌

When washing, pat dry the enzyme-linked immunosorbent assay (ELISA) plate to avoid direct contact between the absorbent paper and the hole. ‌

The color reaction should be avoided from light and the reading should be taken within 30 minutes after termination.

Note: The above is for reference only and is not considered as actual data. Experiments must strictly follow the instructions or consult technical teachers.