Human EGF ELISA development kit contains the key components required for the quantitative measurement of natural and/or recombinant EGF in a sandwich ELISA format within the range of 8-700 pg/mL. Using the ELISA protocol described below, this kit provides sufficient reagents to assay EGF in approximately 1,500 ELISA plate wells.
Contents of Kit
1. PBS: Dilute 10xPBS to 1xPBS, pH 7.2 in sterile water.
2. Wash Buffer: 0.05% Tween-20 in PBS
3. Block Buffer: 1.0% BSA in PBS
4. Diluent: 1.0% BSA in PBS
5. ELISA microplates (Thermo Fisher Cat. # 456529);
6. BSA (Sigma Cat. # A-7030);
7. Stop Solution 2 M Sulfuric Acid (Sigma Cat. # 339741);
8. Dulbecco's PBS [10x] (Gibco BRL Cat. # 14200-075).
Unopened Kit Store at 2-8°C. For more detailed information, please download the document on our website.
Epidermal growth factor or EGF is a growth factor that plays an important role in the regulation of cell growth, proliferation, and differentiation by binding to its receptor EGFR. Human EGF is a 6045-Da protein with 53 amino acid residues and three intramolecular disulfide bonds.
Reconstitution And Storage
Capture Antibody: 720 μg of antigen-affinity purified goat anti-EGF. Centrifuge vial prior to opening. Reconstitute in 1.0 mL PBS for a concentration of 720 μg/mL. Following reconstitution the Capture antibodies may be stored at 2-8°C for up to 6 months. For long term storage it is recommended to aliquot into working volumes and store at -70°C in a manual defrost freezer.
Detection Antibody: 9 μg of biotinylated antigen-affinity purified goat anti-EGF. Centrifuge vial prior to opening. Reconstitute in 1.0 mL sterile 1.0% BSA in PBS for a concentration of 9 μg/mL. Following reconstitution the Detection antibodies may be stored at 2-8°C for up to 6 months. For long term storage it is recommended to aliquot into working volumes and store at -70°C in a manual defrost freezer.
Human EGF Standard: 35 ng of recombinant EGF. Centrifuge vial prior to opening. Reconstitute in 0.5 mL sterile 1.0% BSA in PBS for a concentration of 70 ng/mL. The Standard may be stored at 2-8°C for one month or aliquoted and stored at -70°C for up to three months in a manual defrost freezer.
UltraAvidin-HRP Conjugate: 40 μL vial. Upon receipt, UltraAvidin-HRP conjugate should be stored at 2-8°C, DO NOT FREEZE.
TMB Liquid Substrate: Aspirate and wash plate 4 times. Add 100 μL of TMB HRPO Microwell Substrate Standard Kinetic One Component "Ready Use" to each well. Incubate at room temperature and monitorcolor development with an ELISA plate reader at 450 nm with wavelength correction set at 540 nm or 570 nm. The reaction may be stopped after 15-20 minutes by adding 100 μL of 2 M sulfuric acid to each well.