TPO Antibody – Monoclonal

ELISA: This monoclonal antibody containing culture supernatant has been found by indirect ELISA to bind to recombinant hTPO and hTPO(244-277). Titration experiments show the titer to greater than 1:80 by indirect ELISA

Western Blot: Western immunoblots resulted in a single band being detected at ~ 46 kDa at a dilution of 1:20

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 2% normal goat serum in TBS/Tween-20 buffer.
2. Wash x 2 with TBS/Tween-20.
3. Apply the culture supernatant after dilution at least 1:20. (Note: higher dilutions may be needed) Use 2% normal goat serum in TBS/Tween-20 as buffer for the primary antibody. 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 2% normal goat serum in TBS/Tween-20 buffer. Incubate 1-2 hours. Note: greater sensitivity may be achieved using ABC techniques.
6. Wash x 4 for 5 min. per wash cycle in a fairly large volume of TBS/Tween-20.
7. Develop color using the enhanced DAB reaction.