Human Alpha-fetoprotein, AFP ELISA Kit

Catalog No.: BZEK2322

Size: 96T

Range: 1 μg/L - 16 μg/L

Sensitivity: 0.1 μg/L

Storage and Expiration: Store at 2-8℃ for 6 months. 

Application: For quantitative detection of AFP in human serum, plasma, tissue homogenate or cell culture supernatant.

Introduction

Alpha-fetoprotein(AFP) is a major plasma protein in the fetus, where it is produced by the yolk sac and liver, It is encoded by the AFP gene. The AFP gene is located on chromosome 4q25. AFP is a glycoprotein of 591 amino acids and a carbohydrate moiety. In rodent fetuses AFP binds estradiol to prevent the transport of this hormone across the placenta. The main function of this is to prevent the masculinization of female fetuses.

Principle of the Assay  

This kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. The purified anti- AFP antibody was pre-coated onto 96-well plates. And the HRP conjugated anti- AFP antibody was used as detection antibodies. The standards, test samples and HRP conjugated detection antibody were added to the wells subsequently, mixed and incubated, then, unbound conjugates were washed away with wash buffer. TMB substrates (A & B) were used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the AFP amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of AFP can be calculated.

Kit components

1. One 96-well plate pre-coated with anti- human AFP antibody

2. Standard: 0.5ml (32 μg/L)

3. Standard diluent buffer: 1.5 ml

4. Wash buffer (30×): 20 ml. Dilution: 1:30

5. Sample diluent buffer: 6 ml

6. HRP conjugated anti- human AFP antibody (RTU): 6ml

7. Stop solution: 6 ml

8. TMB substrate A:  6ml

9. TMB substrate B:  6ml

10. Plate sealer:   2

11. Hermetic bag:  1

Material Required But Not Provided

1. 37℃ incubator
2. Microplate reader (wavelength: 450nm)
3. Precise pipette and disposable pipette tips
4. Automated plate washer
5. ELISA shaker
6. 1.5ml of Eppendorf tubes
7. Absorbent filter papers
8. Plastic or glass container with volume of above 1L