title: [Interactions between Hemoglobin and Other Proteins were Proved by the Electrophoresis Release Test and Co-immunoprecipitation].
abstract: OBJECTIVE
  To study the interactions between hemoglobin (Hb) and other proteins within human erythrocytes by using the electrophoresis release test (ERT) and co-immunoprecipitation.
  
  
  METHODS
  First, the fresh normal adult anti-coagulated whole blood was washed to prepare packed RBCs, which were further prepared to erythrocyte suspension and hemolysate.The erythrocyte suspension and hemolysate were analyzed by the electrophoresis release test (ERT) at the same time, and then the band of HbA of erythrocyte sample (RA) and the corresponding band of hemolysate sample (HA) were cut out from the gel and were enriched by freeze-thaw method. Then, the samples were bound with hemoglobin β antibody (37-8) AC, the complexes were separated through 5%-12% SDS-PAGE followed by Q-TOF.
  
  
  RESULTS
  Five bands were found in the gel, each of which was treated by hemoglobin β antibody (37-8) AC, the protein bands of 16,20,22,28 and 50 kD were emerged in RA, HA and RBC lysis.The bands were identified by MS, and the results showed that these bands were hemoglobin, band 3, peroxiredoxin2 (Prx2), band 3 and β-actin, band 3 respectively.
  
  
  CONCLUSIONS
  HbA may interact with Prx2, Band 3 and β-actin, and then the complexes are formed with each other within erythrocytes.
authors.affiliation: Department of Biochemistry and Molecular Biology, Baotou Medical College, Baotou 014060, Inner Mongolia Autonomous Region, China.|Department of Clinical laboratory Examination, Third Affiliated Hospital of Baotou Medical College, Baotou 014030, Inner Mongolia Autonomous Region, China.
authors.forename: Liang
authors.initials: L
authors.lastname: An
authors.affiliation: Department of Biochemistry and Molecular Biology, Baotou Medical College, Baotou 014060, Inner Mongolia Autonomous Region, China.
authors.forename: Xiao-Jing
authors.initials: XJ
authors.lastname: Li
authors.affiliation: Department of Biochemistry and Molecular Biology, Baotou Medical College, Baotou 014060, Inner Mongolia Autonomous Region, China.
authors.forename: Chun-Hua
authors.initials: CH
authors.lastname: Wei
authors.affiliation: Department of Biochemistry and Molecular Biology, Baotou Medical College, Baotou 014060, Inner Mongolia Autonomous Region, China.
authors.forename: Cai-Xia
authors.initials: CX
authors.lastname: Xie
authors.affiliation: Department of Clinical laboratory Examination, Third Affiliated Hospital of Baotou Medical College, Baotou 014030, Inner Mongolia Autonomous Region, China.
authors.forename: Xiao-Hong
authors.initials: XH
authors.lastname: Du
authors.affiliation: Department of Biochemistry and Molecular Biology, Baotou Medical College, Baotou 014060, Inner Mongolia Autonomous Region, China.
authors.forename: Jia-Xin
authors.initials: JX
authors.lastname: Li
authors.affiliation: Department of Biochemistry and Molecular Biology, Baotou Medical College, Baotou 014060, Inner Mongolia Autonomous Region, China. E-mail: synmg@126.com.
authors.forename: Yan
authors.initials: Y
authors.lastname: Su
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes; D006454:Hemoglobins; D006801:Humans; D047468:Immunoprecipitation; D011506:Proteins
chemical_list: D006454:Hemoglobins; D011506:Proteins