Visualization of multiple protein bands on the same nitrocellulose membrane by double immunoblotting.
A method has been developed which allows the simultaneous immunodetection of more than one type of protein on the same nitrocellulose membrane. This procedure does not require special labeling of samples or elution of antibodies from the membrane as do the alternatives cited in the literature (1,2). Proteins are separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electrophoretically transferred to the membrane before specific immunostaining with either peroxidase/4-chloro-1-naphthol or immunogold/silver staining. Antigen identity is visually determined by the formation of different-colored precipitates on the membrane. This innovation in protein blotting offers a savings in time and reagents as well as permitting identification of closely spaced bands with certainty.
Poor
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ML
Lilly Research Laboratories, Eli Lilly & Company, Indianapolis, Indiana 46285.
Santa
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PF
Sittampalam
G S
GS
D001426:Bacterial Proteins / Q000302:isolation & purification; D015153:Blotting, Western / Q000379:methods*; D003101:Collodion; D006046:Gold; D013006:Growth Hormone / Q000031:analogs & derivatives / Q000302:isolation & purification; D006735:Horseradish Peroxidase; D019382:Human Growth Hormone; D009284:Naphthols; D011506:Proteins / Q000302:isolation & purification*; D012834:Silver; D013194:Staining and Labeling
D001426:Bacterial Proteins; D009284:Naphthols; D011506:Proteins; D019382:Human Growth Hormone; D012834:Silver; C025917:4-chloro-1-naphthol; D006046:Gold; D013006:Growth Hormone; D003101:Collodion; D006735:Horseradish Peroxidase