IGF Antibody – Monoclonal

ELISA: This IgG fraction has been found by indirect ELISA to bind to hIGF-1 at concentrations of 50 ng/well or less.

Western Blot: Western immunoblots resulted in a single band being detected at ~ 7.5 kDa at a dilution of 1:5000

Western Blot Protocol: 1. After SDS-PAGE (on either 4-15% gradient gels or single percentage gels. such at 12% gels) and electrophoretic transfer to PVDF membrane. block the membrane overnight with 4% normal goat serum in TBS/Tween-20 buffer.
2. Wash x 2 with TBS/Tween-20.
3. Apply the monoclonal antibody after dilution at least 25 ng/ml. (Note: higher dilutions may be needed). Use 1% normal goat serum in TBS/Tween-20 as buffer. Let the primary antibody bind for 2-4 hours.
4. Wash x 3 with TBS/Tween-20.
5. Apply affinity purified HRP-goat anti-mouse IgG antiserum diluted 1:2500 (Note: dilution may vary depending upon supplier) in 1% normal goat serum in TBS/Tween-20. Incubate 1-2 hours. Note: greater sensitivity may be achieved using ABC techniques.
6. Wash x 4 for 5 minutes per cycle in TBS/Tween-20.
7. Develop color using the enhanced DAB reaction or develop film using ECL or other chemiluminescent techniques.