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 mouse 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.
Range: 15.6 pg/ml-1000 pg/ml
Sensitivity < 1 pg/ml
Storage and Expiration: Store at 2-8℃ for 4 months, or at -20℃ for 8 months.
Application: For quantitative detection of EGF in mouse serum, plasama, body fluids, tissue lysates or cell culture supernate.
1. One 96-well plate pre-coated with anti-Mouse EGF antibody
2. Lyophilized Mouse EGF standards: 2 tubes (10 ng / tube)
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-Mouse 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.
Swiss Port: P01132
Gene ID: 13645
Please check datasheet for protocol.