A nonradioactive dot blot assay for transglutaminase activity.
Aberrant transglutaminase (TG) activity has been implicated in the pathology of numerous diseases, including Huntington's disease and Alzheimer's disease. To fully characterize the role of TGs in these disorders, it is important that simple quantifiable assays be made available. The most commonly used assay currently employed requires significant time and a radioactive substrate. The assay described here uses a biotinylated substrate in conjunction with a dot blot apparatus to eliminate the use of radioactive substrates and allows relative transglutaminase activity to be measured simultaneously with minimal sample preparation in a large number of samples containing purified enzyme, cell extracts, or tissue homogenates.
McConoughey
Stephen J
SJ
Burke Medical Research Institute, White Plains, NY 10605, USA. mcconoughey.4@osu.edu
Niatsetskaya
Zoya V
ZV
Pasternack
Ralf
R
Hils
Martin
M
Ratan
Rajiv R
RR
Cooper
Arthur J L
AJ
D000588:Amines / Q000737:chemistry; D000818:Animals; D001710:Biotin / Q000031:analogs & derivatives / Q000737:chemistry; D019921:Biotinylation; D003720:Densitometry; D019204:GTP-Binding Proteins / Q000032:analysis / Q000378:metabolism; D006168:Guinea Pigs; D008163:Luminescent Measurements / Q000379:methods*; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011503:Transglutaminases / Q000032:analysis* / Q000378:metabolism
D000588:Amines; C076022:5-(biotinamido)pentylamine; D001710:Biotin; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011503:Transglutaminases; D019204:GTP-Binding Proteins