Improvements of Western blotting to detect monoclonal antibodies.
A comparison of the effects of different factors on the sensitivity of Western blotting technique to detect monoclonal antibodies is described. The major improvements were obtained by: A) renaturating the antigen in the gel before transferring it in carbonate buffer at pH 10 onto nitrocellulose and B) using alkaline-phosphatase-conjugated second antibody instead of peroxidase-conjugated second antibody.
Bestagno
M
M
Cerino
A
A
Riva
S
S
Astaldi Ricotti
G C
GC
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis*; D006367:HeLa Cells / Q000032:analysis; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D006801:Humans; D007124:Immunoenzyme Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D008832:Microchemistry; D011489:Protein Denaturation; D012261:Ribonucleoproteins / Q000032:analysis
D000911:Antibodies, Monoclonal; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D012261:Ribonucleoproteins