Hi all, I want to get suggestion from anyone with regards to this situation. We tried to extract protein from coral tissue (5cm fragment length; airbrushed the fragment and homogenised) in the lab, and the we got very low protein concentration yield (0.01-0.1 ug/ug). The lysis buffer used; 0.01M Tris-HCl, 0.5M sucrose, 0.001M EDTA, 0.15M KCl and 0.001M PMSF. We plan to do 2DE gel for later analysis.
Does anyone think the buffer require modification?
Thank you in advance!
Mohsen Miandehy thank you so much for sharing. will definitely go through the protocol!
Downs, C.A. (2005) Cellular diagnostics and its application to aquatic and marine toxicology. In: G. Ostrander (ed) Techniques in aquatic toxicology. 2. CRC Press, Inc., Boca Raton, pp 301‐313 Ghosh, S., Gepstein, S., Heikkila, J.J., et al. (1988) Use of a scanning densitometer or an ELISA plate reader for measurement of nanogram amounts of protein in crude extracts from biological tissues. Analytical Biochemistry, 169, 227‐233. Rasband, W.S. (2011) ImageJ, U. S. National Institutes of Health, Bethesda, Maryland, USA, http://imagej.nih.gov/ij/, 1997‐2011. Sokal, R.R. and Rohlf, F.J. (1995) Linear Regression. In: Biometry the principles and practice of statistics in biological research, 3rd edn. pp. 452‐455. W.H. Freeman & Co., New York. -- can you consult these please?
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