Bio-Rad Trans-Blot® SD Semi-Dry Transfer Cell - Manual (Page 18)

8.4 Poor Detection Sensitivity or No Reactivity
1. Consult detection kit manual.
2. Antigen binding is incomplete. See Troubleshooting Sections 8.1–8.3.
3. Antibody reaction times are insufficient. Increase reaction times.
4. Sample load is insufficient. Increase the protein concentration applied to the gel.
5. Antigen may require specific temperature regulation during transfer to prevent
denaturation. Use the Trans-Blot cell with the super cooling coil to transfer heat-sensitive
proteins.
6. Monoclonal antibodies might not recognize a denatured antigen. Assess binding of other
monoclonals or polyclonal antibodies. Blot native proteins.
Section 9
References
1. Southern, E. M., J. Mol. Biol., 98, 503 (1975).
2. Alwine, J. C., Kemp, D. J., Parker, B. A., Reiser, J., Stark, G. R. and Wahl, G. W., Methods
Enzymol., 68, 220 (1979).
3. Thomas, P. S., PNAS, 77, 5201 (1980).
4. Seed, B., Nuc. Acids Res., 10, 1799 (1982).
5. Renart, J., Peiser, J. and Stark, G. R., PNAS, 76, 3116 (1979).
6. Bowen, P., Steinberg, J., Laemmli, U. K. and Weintraub, H., Nuc. Acids Res., 8 (1980).
7. Towbin, H., Staehelin, T. and Gordon, J., PNAS, 76, 4350 (1979).
8. Bittner, M., Kupferer, P. and Morris, C. F., Anal. Biochem., 102, 459 (1980).
9. Stellwag, E. J. and Dahlberg, A. E., Nuc. Acids Res., 8, 299 (1980).
10. Kutateladze, T. V., Axelrod, V. D., Gorbulev, V. G., Belzhelarskya, S. N. and Vartikyan, R. M.,
Anal. Biochem., 100, 129 (1979).
11. Peudelhuber, T. L., Ball, D. J., Davis, A. H. and Garrad, W. J., Nuc. Acids Res., 10, 1311 (1982).
12. Danner, D. B., Anal. Biochem., 125, 139 (1982).
13. Bio-Rad Technical Bulletin 1110 “Zeta-Probe Blotting Membranes” (1987).
14. Holland, L. J. and Wangh, L. J., Nuc. Acids Res., 10, 3282, (1983).
15. Khyse-Andersen, J., Biochem. Biophys. Meth., 10, 203, (1984).
16. Bjerrum, O. J. and Schafer-Nielsen, C., Analytical Electrophoresis, M. J. Dunn, ed., p. 315; Verlag
Chemie, Weinheim, (1986).
17. Dunn, S. D., Anal. Biochem., 157, 144 (1986).
18. Bio-Rad Laboratories, Zeta-Probe Instruction Manual (1986).
19. Anderson, N. L., Nance, S. L., Pearson, T. W. and Anderson, N. G., Electrophoresis, 3, 135 (1982).
20. Howe, J. G. and Hershey, J. W. B., J. Biol. Chem., 256, 12836 (1981).
21. Erickson, P. F., Minier, L. N. and Lasher, P. S., J. Immun. Meth., 51, 241 (1982).
22. Polvino, W. J., Saravis, C. A., Sampson, C. E. and Cook, R. B., Electrophoresis, 4, 368 (1983).
23. Tovey, E. and Baldo, B. A., Electrophoresis, 8, 384 (1987).
24. Gershoni, J. M. and Palade, G. E., Anal, Biochem., 131, 1 (1983).
25. Gershoni, J. M. and Palade, G. E., Anal. Biochem., 124, 396 (1982).
26. Bio-Rad Laboratories, Biotin-Blot Total Protein Stain Instruction Manual (1985).
27. LaRochelle, W. J. and Froehner, S. C., J. Immun. Meth., 92, 65 (1986).
28. Johnson, D. A., Gautsch, J. W., Sportsman, J. R. and Elder, J. H., Gene Anal. Tech., 1, 3 (1984).
*Spectr/Por is a trademark of Spectrum Medical Industries.
*Coomassie is a trademark of I C I Organics, Inc.
15