Polyacrylamide gel electrophoresis and immunoblotting of proteins extracted from paraffin-embedded tissue sections.
We show in this communication that polyacrylamide gel electrophoresis (PAGE) and immunoblotting of proteins can be performed using one to two 5-7 micron paraffin sections of tissues fixed in non-cross-linking fixatives (acetone, alcohol, or modified Carnoy's solution). Proteins for study were extracted from paraffin sections of mouse foot pad and liver. The presence of unaltered keratin polypeptides in tissues fixed with either acetone or alcohol was demonstrated in gels stained with Coomassie brilliant blue. The preservation of their antigenic determinants was demonstrated with immunoblotting. Furthermore, the immunoreactivity of soluble proteins, such as albumin, remained unaltered in immunoblots obtained from paraffin-embedded mouse liver sections. These data indicate that tissues embedded and stored in paraffin are useful for the above-mentioned biochemical and immunological studies and may therefore be an important technique for diagnostic pathology or retrospective studies.
Conti
C J
CJ
University of Texas System Cancer Center, Science Park-Research Division, Smithville 78957.
Larcher
F
F
Chesner
J
J
Aldaz
C M
CM
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D007163:Immunosorbent Techniques; D051379:Mice; D008970:Molecular Weight; D010232:Paraffin; D011506:Proteins / Q000032:analysis*; D012394:Rosaniline Dyes
D011506:Proteins; D012394:Rosaniline Dyes; D010232:Paraffin; C004692:coomassie Brilliant Blue