Western blot analysis of Human EGF was performed by loading 2ug of recombinant human EGF (Lane 1) or supernatant from an EGF expression clone transfected in 293T cells onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% Milk/TBST for at least 1 hour. Membranes were probed with an EGF monoclonal antibody recognizing Human EGF (Product # M806) at a dilution of 1:5000 overnight at 4°C on a rocking platform. Membranes were washed in TBS-0.1%Tween 20 and probed with a goat anti-rabbit-HRP secondary antibody (Product # 31430) at a dilution of 1:10,000 for at least one hour. Membranes were washed and chemiluminescent detection performed using Super Signal West Dura (Product # 34075).
|Tested species reactivity||Human|
|Host / Isotype||Mouse / IgG|
|Immunogen||Recombinant Human EGF|
|Storage buffer||PBS with 4% BSA|
|Tested Applications||Dilution *|
|ELISA (ELISA)||0.125 - 0.25 µg/ml|
|Western Blot (WB)||1:1000 - 1:5000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
The M806B anti-EGF antibody (biotinylated conjugate of clone 3A8) has successfully been paired as the detection antibody in a sandwich ELISA with coating antibody M805 (Clone 1H11).
Typical dilutions for sandwich ELISA: Coat = 1-3 µg/ml and Detection = 0.125-0.5 µg/ml.
The protein encoded by EGF gene is a growth factor that stimulates 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
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.