Large-scale chromatofocusing-based method for isolating thymosin beta 4 and thymosin beta 9 from bovine tissues.
A large-scale method for the isolation of thymosin beta 4 (up to 120 mg) and thymosin beta 9 (up to 40 mg) from bovine lung (up to 2 kg) was developed. The isolation protocol included tissue homogenization in 0.4 M HClO4, centrifugation, solid-phase extraction through LiChroprep RP-18 material, chromatofocusing on polybuffer exchanger PBE 94-modified Sepharose and dialysis against water. The isolated products were characterized by analytical isoelectric focusing, reversed-phase HPLC, electrospray ionization mass spectrometry and amino acid analysis. The method developed is rapid and convenient, requires no expensive equipment and can be used for the isolation of thymosin beta 4 and homologous peptides from various animal tissues.
Roboti
A
A
National Centre for Scientific Research Demokritos, Institute of Radioisotopes/Radiodiagnostic Products, Aghia Paraskevi Attikis, Athens, Greece.
Livaniou
E
E
Evangelatos
G P
GP
Tsoupras
G
G
Tsolas
O
O
Ithakissios
D S
DS
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D004797:Enzyme-Linked Immunosorbent Assay; D007202:Indicators and Reagents; D007525:Isoelectric Focusing; D008168:Lung / Q000737:chemistry; D013058:Mass Spectrometry; D008840:Microfilament Proteins / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D013947:Thymosin / Q000302:isolation & purification*; D000099201:Thymosin beta(4)
D007202:Indicators and Reagents; D008840:Microfilament Proteins; D013947:Thymosin; D000099201:Thymosin beta(4); C042362:thymosin beta(9)