* Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own experiment using appropriate negative and positive controls.
Product Specific Information
The M807 anti-EGF antibody has successfully been paired as the detection antibody in a sandwich ELISA with coating antibody MA805 (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.
EGF Antibody (M807) in WB M807_WB_Image.jpg
Western Blot with anti-EGF Monoclonal Antibody [9A1] (M807)
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 # M807) 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).
EGF Antibody (M807) in IF EGF_Monoclonal_Antibody_M807_Immunofluorescence_1.jpg
Immunofluorescence with anti-EGF Monoclonal Antibody [9A1] (M807)
Immunofluorescent analysis of EGF (green) in HeLa cells either left serum starved (left panel) or treated with serum (right panel) for 30 minutes. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with an EGF monoclonal antibody (Product # M807) at a dilution of 1:50 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-mouse IgG secondary antibody (Product # 35502) at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight 554 Phalloidin (Product # 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan at 20X magnification.
(This product is for In Vitro experimental use only. Not for resale without express authorization.)