Technical Manual for Human Interleukin-17 (IL-17) ELISA Detection Kit

BS-9770 Human IL-17 ELISA Kit

1、 Product Overview

This kit uses a double antibody sandwich enzyme-linked immunosorbent assay (ELISA) method, specifically designed for quantitative detection of interleukin-17 (IL-17) levels in human serum, plasma, and related biological fluid samples. IL-17 is a pro-inflammatory cytokine secreted by Th17 cells that plays a critical role in autoimmune diseases and inflammatory responses.

2、 Detection principle

Double antibody sandwich technology:

Pre packaged microplates with anti human IL-17 monoclonal antibodies capture IL-17 in the sample.

Add horseradish peroxidase (HRP) labeled secondary antibody to form an "antibody antigen enzyme labeled antibody" complex.

Through TMB colorimetric reaction, the color depth is positively correlated with IL-17 concentration, and the absorbance (OD value) is measured at 450nm wavelength.

3、 Composition of reagent kit

Component specifications (96 well configuration) Storage conditions

Enzyme labeled coated plate 1 × 96 well 2-8 ℃

Standard product (270ng/L) 0.5ml x 1 bottle 2-8 ℃ (long-term -20 ℃)

Standard diluent 1.5ml x 1 bottle 2-8 ℃

Enzyme labeled reagent 6ml x 1 bottle 2-8 ℃

Sample diluent 6ml x 1 bottle 2-8 ℃

6ml each of color reagent A/B solution in 1 bottle, 2-8 ℃, away from light

Termination solution 6ml x 1 bottle 2-8 ℃

Concentrated detergent 20ml x 30x x 1 bottle 2-8 ℃

1 copy of manual at room temperature

4、 Sample processing requirements

Serum/plasma:

Let it stand at room temperature for 10-20 minutes, rotate at 2000-3000 rpm/centrifuge for 20 minutes, and collect the supernatant

Avoid repeated freeze-thaw cycles. If sedimentation occurs, centrifuge again

Cell culture supernatant:

After centrifugation, sterile operation is required to collect the supernatant and detect secretory components

Urine/tissue homogenate:

Centrifuge the urine and collect the supernatant; The tissue sample needs to be homogenized and centrifuged for packaging

5、 Operation steps

Standard dilution:

Gradient dilution standard (such as 270ng/L to 1ng/L), add 50 μ L of sample per well

Sample addition:

Dilute the sample and add 10 μ L to the micropores, with a sample volume of 40 μ L (total dilution of 5 times)

Nurturing:

React at 37 ℃ in the dark for 30 minutes, covered with a sealing film

wash

Soak in detergent for 1 minute/time, repeat 3-5 times, and pat dry the liquid in the hole.

Color rendering and termination:

Add TMB color solution and react in the dark for 15-20 minutes, then stop the reaction with the stop solution.

reading:

Measure the OD value at a wavelength of 450nm using an enzyme-linked immunosorbent assay (ELISA) reader, and calculate the concentration using a standard curve.

6、 Performance indicators

Sensitivity: as low as 0.1 pg/mL (mouse IL-17 reference value), human IL-17 is approximately 3.8 pg/mL.

Detection range: 7.8-500 pg/mL (reference value for human IL-17C), or 31.3-2000 pg/mL (human IL-17).

Specificity: No cross reactivity with other members of the IL-17 family, such as IL-17B and IL-17F.

Repeatability: coefficient of variation within/between plates<10%

7、 Precautions

Storage conditions:

The reagent kit should be stored at 2-8 ℃, and standard samples should be stored at -20 ℃ for long-term use

Validity period:

6 months (4 ℃), 12 months (-20 ℃).

Experimental environment:

Avoid strong light exposure, and the color developing solution needs to be pre equilibrated at 37 ℃.

8、 References

Principle and operating specifications of double antibody sandwich method

Guidelines for Sample Processing and Preservation

Test kit performance validation data

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

Tianjin Bensheng has always regarded quality control as the lifeblood of the enterprise and pursued continuous improvement of its competitiveness in the field of human interleukin-17 (IL-17) ELISA detection kit. The company always adheres to the corporate spirit of abiding by laws and regulations, being strict with oneself, being tolerant and willing to take on responsibilities as the standard in its operations. It maintains and expands the market with excellent quality and service, and maximizes the satisfaction of customer needs. Our development goal is to achieve mutual benefit with our customers. This student! A partner you trust. We are willing to cooperate sincerely with you and create a better future together.