University of Alabama at Birmingham
Question
Asked 7th Mar, 2015
What are the disadvantages of activity stain ?
The gel electrophoresis was performed using the 1% agarose, pH=8.6 on lactate dehydrogenase from heart and skeletal muscle samples. The results obtained were quite poor, no clear bands. I was wondering how I could explain this?
Also I know what the advantages of activity stain are, but could you tell me what are possible disadvantages of activity stain (the one used in this experiment was: 100mM sodium lactate 0.2% NAD 0.6% tetrazolium dye (MTT) 0.005% medola blue pH 7.5)
All Answers (1)
I do not have experience with agarose gel or LDH. But I have done lot of activity Native Page gels for Catalase, Isocitrate dehyrogease, MMPs, etc...
Native PAGE provides high resolution of bands by reducing the diffusion of proteins both while running the gels as well as during incubation with the reaction buffer. One of the major disadvantage of activity gels is that it doesn't provide the specific activity of the enzyme. You can only visualize the activity in your sample.
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