title: One-dimensional and two-dimensional immobilized metal affinity electrophoresis.
abstract: Immobilized metal affinity electrophoresis (IMAEP) is a straightforward method in which metal ions are embedded in a polyacrylamide gel strip with a negligible electrophoretic migration. Due to the preferential binding between metal ions and the phosphate group, this method uses immobilized metal ions like iron, manganese, aluminum, or titanium to capture phosphoproteins from a mixture of phosphoprotein and nonphosphoproteins. IMAEP has also been incorporated into a traditional two-dimensional (2D) sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) system (isoelectric focusing-PAGE) to increase its resolving power. In 2D IMAEP, the metal ions in polyacrylamide gel strip are overlaid on top of the second dimensional polyacrylamide gel to stop electrophoretic migration of phosphoproteins. Data shows that there is no detrimental effect of SDS in IMAEP on the extraction of phosphoproteins from a mixture of proteins. In addition, SDS exposes phosphate groups by unfolding the phosphoproteins to facilitate metal ion-phosphate binding while supplying the protein with negative charges.
authors.affiliation: University of Illinois at Chicago, Chicago, IL, USA. boblee@uic.edu
authors.forename: Bao-Shiang
authors.initials: BS
authors.lastname: Lee
authors.forename: G D
authors.initials: GD
authors.lastname: Lasanthi
authors.forename: P
authors.initials: P
authors.lastname: Jayathilaka
authors.forename: Jin-Sheng
authors.initials: JS
authors.lastname: Huang
authors.forename: Shalini
authors.initials: S
authors.lastname: Gupta
mesh_terms: D002021:Buffers; D002256:Carbonic Anhydrases / Q000737:chemistry / Q000302:isolation & purification; D002364:Caseins / Q000737:chemistry / Q000302:isolation & purification*; D045744:Cell Line, Tumor; D004396:Coloring Agents / Q000737:chemistry; D056831:Coordination Complexes / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D006454:Hemoglobins / Q000737:chemistry / Q000302:isolation & purification; D006801:Humans; D007526:Isoelectric Point; D008670:Metals / Q000737:chemistry; D008970:Molecular Weight; D011485:Protein Binding; D012394:Rosaniline Dyes / Q000737:chemistry; D012709:Serum Albumin / Q000737:chemistry / Q000302:isolation & purification; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D002021:Buffers; D002364:Caseins; D004396:Coloring Agents; D056831:Coordination Complexes; D006454:Hemoglobins; D008670:Metals; D012394:Rosaniline Dyes; D012709:Serum Albumin; D002256:Carbonic Anhydrases; C004692:coomassie Brilliant Blue