Direct tissue isoelectric focusing on ultrathin polyacrylamide gels. Applications in enzyme, lectin and immunohistochemistry.
Application of cryostat sections directly onto ultrathin polyacrylamide gels and subsequent isoelectric focusing allows elution of proteins, glycoproteins and peptides out of the sections into the gels. The eluted compounds reveal clearly delineated band patterns in the polyacrylamide gels. The advantage of this method is that enzyme histochemical reactions can be directly performed in the gel and in the electroeluted tissue sections. Therefore, this method is suitable for specifying, in more detail, histochemical enzyme reactions and for detecting multiple forms of enzymes even from a single tissue section. Furthermore, the transfer of proteins, glycoproteins and peptides from the gel onto nitrocellulose by a modified Western blot procedure offers the possibility of checking findings obtained by lectin histochemistry and immunohistochemistry.
Ludwig-Maximilians-Universität München, Germany.
U
U
Schumacher
P
P
Trudrung
M
M
Ruhnke
R
R
Gossrau
D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000166:cytology / Q000378:metabolism; D003101:Collodion; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D004798:Enzymes / Q000378:metabolism*; D006023:Glycoproteins / Q000378:metabolism; D007150:Immunohistochemistry / Q000379:methods; D007421:Intestine, Small / Q000166:cytology / Q000378:metabolism; D007525:Isoelectric Focusing / Q000379:methods*; D007668:Kidney / Q000166:cytology / Q000378:metabolism; D037102:Lectins / Q000378:metabolism*; D008099:Liver / Q000166:cytology / Q000378:metabolism; D009206:Myocardium / Q000166:cytology / Q000378:metabolism; D009479:Neuropeptides / Q000378:metabolism; D051381:Rats; D013154:Spleen / Q000166:cytology / Q000378:metabolism; D013194:Staining and Labeling / Q000379:methods
D004798:Enzymes; D006023:Glycoproteins; D037102:Lectins; D009479:Neuropeptides; D003101:Collodion