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Position: Discussion forums > Bioscience biotechniques discussion forums > Proteomics and protein biochemistry forum
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protein estimation problem IMP
Posted by: kum81 (IP Hidden, Junior member, 10)
Date: September 19, 2005 04:14AM
hi
i perform whole cell protein profiling of bacteria. the cell pellet (~30mg) is denatured by dissolving in 80 microlt of 1X SDS-loading dye (containing BBB) and boiled for 5 min. loading 10 microlitres in a 10%SDS PAGE (10cm x 7 cm x .1cm) gives a fairly good profile. i want to do protein estimation of this sample. i tried lowry's and bradford method but the amount obtained is very very less than that would be sufficient to show such good profile. if protein extracted by sonication then 40micrograms loading gives the similar profile. can someone suggest a suitable protein estimation method. thanx
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