Rat EGF ELISA Kit

Catalog No.: BEK1282

Size: 96T

Range: 7.8 pg/ml - 500 pg/ml

Sensitivity < 1 pg/ml

Storage and Expiration: Store at 2-8℃ for 6 months, or at -20℃ for 12 months.

Application: For quantitative detection of EGF in rat serum, plasama, body fluids, tissue lysates or cell culture supernate.

Introduction

Epidermal growth factor (EGF) is a growth factor that stimulates cell growth, proliferation, and differentiation by binding to its receptor EGFR. It has a profound effect on the differentiation of specific cells in vivo and is a potent mitogenic factor for a variety of cultured cells of both ectodermal and mesodermal origin. EGF is produced in abundance by the rat submandibular gland, and it can be found in human platelets, macrophages, urine, saliva, milk, and plasma. It is a magnesiotropic hormone crucial for total body Mg(2+) balance. Futamura et al. (2002) found that chronic treatment of rats with haloperidol had no influence on EGF levels in the brain or serum.  

Principle of the Assay 

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-EGF polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-EGF polyclonal antibody was used as detection antibodies. The standards, test samples and biotin conjugated detection antibody were added to the wells subsequently, and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates 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 EGF amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of EGF can be calculated.

Kit components

1. One 96-well plate pre-coated with anti-rat EGF antibody
2. Lyophilized rat EGF standards: 2 tubes (10ng / tube)
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-rat EGF antibody (Concentrated): 130μl. Dilution: 1:100
5. Antibody diluent buffer: 12ml
6. Avidin-Biotin-Peroxidase Complex (ABC) (Concentrated): 130μl. Dilution: 1:100
7. ABC diluent buffer: 12ml
8. TMB substrate: 10ml
9. Stop solution: 10ml
10. Wash buffer (25X): 30ml

Note: Reconstitute standards and test samples with Kit Component 3.

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. Plate cover
8. Absorbent filter papers
9. Plastic or glass container with volume of above 1L