title: Optimal protein extraction methods from diverse sample types for protein profiling by using Two-Dimensional Electrophoresis (2DE).
abstract: There is a great diversity of protein samples types and origins, therefore the   optimal procedure for each sample type must be determined empirically. In order to obtain a   reproducible and complete sample presentation which view as many proteins as possible on   the desired 2DE gel, it is critical to perform additional sample preparation steps to improve   the quality of the final results, yet without selectively losing the proteins. To address this, we   developed a general method that is suitable for diverse sample types based on phenolchloroform   extraction method (represented by TRI reagent). This method was found to yield   good results when used to analyze human breast cancer cell line (MCF-7), Vibrio cholerae,   Cryptocaryon irritans cyst and liver abscess fat tissue. These types represent cell line,   bacteria, parasite cyst and pus respectively. For each type of samples, several attempts were   made to methodically compare protein isolation methods using TRI-reagent Kit, EasyBlue Kit,   PRO-PREP™ Protein Extraction Solution and lysis buffer. The most useful protocol allows the   extraction and separation of a wide diversity of protein samples that is reproducible among   repeated experiments. Our results demonstrated that the modified TRI-reagent Kit had the   highest protein yield as well as the greatest number of total proteins spots count for all type   of samples. Distinctive differences in spot patterns were also observed in the 2DE gel of   different extraction methods used for each type of sample.
authors.affiliation: Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia.
authors.forename: A A
authors.initials: AA
authors.lastname: Tan
authors.forename: S N
authors.initials: SN
authors.lastname: Azman
authors.forename: N R
authors.initials: NR
authors.lastname: Abdul Rani
authors.forename: B C
authors.initials: BC
authors.lastname: Kua
authors.forename: S
authors.initials: S
authors.lastname: Sasidharan
authors.forename: L V
authors.initials: LV
authors.lastname: Kiew
authors.forename: N
authors.initials: N
authors.lastname: Othman
authors.forename: R
authors.initials: R
authors.lastname: Noordin
authors.forename: Y
authors.initials: Y
authors.lastname: Chen
mesh_terms: D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D008967:Molecular Biology / Q000379:methods*; D011506:Proteins / Q000032:analysis* / Q000302:isolation & purification*; D020543:Proteome / Q000032:analysis*
chemical_list: D011506:Proteins; D020543:Proteome