TY - JOUR
T1 - A new electrophoretic-autoradiographic method for the visual detection of phosphotransferases
AU - Tischfield, Jay A.
AU - Bernhard, H. P.
AU - Ruddle, Frank H.
N1 - Funding Information:
The authors acknowledge the excellent technical aasistancc of Ms. Pauline Chao. This work was supported by NIH Grant GM 09966, and a G. S. Public Health Service Traineeship to one of the authors (J. A. T.).
PY - 1973/6
Y1 - 1973/6
N2 - In this paper we present the details of a slab acrylamide, lanthanum precipitation, autoradiographic technique and show it to be useful for the visualization of at least four different enzymes. We believe that with appropriate separation conditions and reaction mixtures this technique could be extended to a larger number of enzymes, theoretically all those whose isotopically labeled product could be specifically precipitated within the matrix of a polyacrylamide gel. It will be interesting to use this technique with other gel buffer systems, particularly those with a lower pH that have recently been reported (21). In addition, it might be useful to combine it with isoelectric focusing in slab gels (10). The technique would appear to be particularly useful for phosphotransferases and, to date, it has been applied to thymidine kinase and adenosine kinase with encouraging results. Work with other enzymes, and adapting the technique to starch gels, is also in progress.
AB - In this paper we present the details of a slab acrylamide, lanthanum precipitation, autoradiographic technique and show it to be useful for the visualization of at least four different enzymes. We believe that with appropriate separation conditions and reaction mixtures this technique could be extended to a larger number of enzymes, theoretically all those whose isotopically labeled product could be specifically precipitated within the matrix of a polyacrylamide gel. It will be interesting to use this technique with other gel buffer systems, particularly those with a lower pH that have recently been reported (21). In addition, it might be useful to combine it with isoelectric focusing in slab gels (10). The technique would appear to be particularly useful for phosphotransferases and, to date, it has been applied to thymidine kinase and adenosine kinase with encouraging results. Work with other enzymes, and adapting the technique to starch gels, is also in progress.
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U2 - 10.1016/0003-2697(73)90105-X
DO - 10.1016/0003-2697(73)90105-X
M3 - Article
C2 - 4351949
AN - SCOPUS:0015829662
SN - 0003-2697
VL - 53
SP - 545
EP - 554
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 2
ER -