Biotechnology and Bioinformatics (1+1)

Protocol

1. Dot 1-2 ul of the test sample (positive antigen) in case antibody detection or the suspected sample suspension in case antigen detection on to a NC or PVDF membrane and air dry.

2. Incubate the membrane in the blocking solution for 1 hr at 37^oC

3. Rinse the membrane for three times in PBST

4. Incubate the membrane in standardized dilution of test antibody (for antibody detection) or the primary antibody for 1 hr at 37^oC

5. Rinse the membrane for three times in PBST for three times 5 minutes each

6. Incubate the membrane in standardized dilution of secondary antibody enzyme conjugate

7. for 1 hr at 37^oC

8. Rinse the membrane for three times in PBST for three times 5 minutes each

9. Expose the membrane to the substrate solution

10. The positive result are visible as a dot