Technical Manual for Porcine Epidermal Growth Factor (EGF) ELISA Detection Kit

BS-4568 Porcine Epidermal Growth Factor (EGF) ELISA Kit

1、 Product Overview

This kit uses double antibody sandwich enzyme-linked immunosorbent assay technology, specifically designed for quantitative detection of epidermal growth factor (EGF) content in pig serum, plasma, tissue homogenate, and cell culture supernatant. The product is for scientific research purposes only and is suitable for fields such as pig growth regulation and disease model research.

2、 Detection principle

Solid phase antibody coating: Pre coating anti porcine EGF monoclonal antibody onto a microporous plate to form a solid phase carrier

Antigen antibody reaction: EGF in the sample binds to solid-phase antibodies, and then biotinylated secondary antibodies are added to form sandwich complexes

Enzyme linked immunosorbent assay (ELISA): HRP labeled streptavidin is added to catalyze the color development of TMB substrate, and the color intensity is positively correlated with the concentration of EGF in the sample

Quantitative analysis: Measure absorbance at 450nm wavelength and calculate concentration through standard curve

3、 Composition of reagent kit

Storage conditions for component specifications

Pre packaged microporous plate with 96 holes/plate sealed at 2-8 ℃

Standard product (freeze-dried) 6 tubes -20 ℃

Enzyme labeled secondary antibody 6ml, 2-8 ℃, away from light

6ml of color developer A/B solution at 2-8 ℃, away from light

Termination solution 6ml at room temperature

20 x 20ml washing buffer at room temperature

Sample diluent 6ml 2-8 ℃

1 copy of instruction manual-

4、 Sample processing

Collection requirements:

Serum/plasma: EDTA anticoagulant, let stand to precipitate serum and centrifuge (3000rpm × 15min)

Tissue homogenate: Grind in physiological saline at a ratio of 1:9, centrifuge to obtain supernatant

Cell supernatant: Centrifuge at 1000rpm for 10 minutes to remove impurities

Storage conditions:

Short term: 2-8 ℃ (≤ 3 days)

Long term: -20 ℃ (avoid repeated freezing and thawing)

5、 Operation steps

Standard dilution:

Prepare 480pg/ml stock solution with standard diluent and dilute it in a 2-fold gradient to 0pg/ml

Dilution sequence: 480 → 240 → 120 → 60 → 30 → 0pg/ml

Sample addition reaction:

Add 50 μ l of standard/sample to each well, and do not add blank wells

Incubate at 37 ℃ for 30 minutes and wash the plate 5 times

Enzyme linked reaction:

Add 50 μ l of enzyme-linked immunosorbent assay secondary antibody and incubate at 37 ℃ for 30 minutes

After washing the board, add TMB color solution and react in the dark for 10 minutes

Termination of measurement:

Add 50 μ l of termination solution and measure the absorbance at 450nm within 30 minutes

6、 Result calculation

Draw a standard curve with standard concentration (pg/ml) as the horizontal axis and OD value as the vertical axis

Sample concentration=standard curve fitting value x sample dilution factor (recommended 5-fold dilution)

7、 Performance parameters

Indicator parameters

Detection range 7.8-500pg/ml

Sensitivity 2.51pg/ml

批内精密度 CV<6.6%

Batch precision CV<7.9%

Recovery rate 91-109%

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

Tianjin Bensheng has always regarded quality control as the lifeblood of the enterprise and pursued continuous improvement of its competitiveness, using the Porcine Epidermal Growth Factor (EGF) 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. Win win cooperation with customers is our development goal. We are your trusted partner!