Immobilized Ficin was specifically designed for cleavage of mouse IgG1 into F(ab')2 or Fab fragments. Ficin will generate mouse IgG1 F(ab')2 and Fab fragments in the presence of 1 mM cysteine and 10 mM cysteine, respectively. Fragment generation from other species and isotypes may also be possible through modification of the cysteine concentration and other digestion parameters.Ficin isolated from fig latex, is a sulfhydryl protease with a similar active site to bromelain and papain. These enzymes vary in kinetics and stereochemical selectivity and can generate F(ab')2 and Fab fragments. However, yields and immunoreactivity are better when digesting mouse IgG1 with ficin. Ficin acts on bonds involving uncharged and/or aromatic amino acids.The immobilization of ficin enhances stability against denaturation, heat and autolysis. Immobilization also eliminates any potential for antibody-enzyme adducts that cause continued sample digestion. In addition to these advantages, immobilization allows the ficin to be reused.
Specific Activity:≥200 Bapa U/g
Extinction Coefficient (E1%):21.0 (280nm)(Lit.)
Appearance:White powder
Isoelectric point (pI):9.0 (Lit.)
pH:5.0-7.5 (optimum pH for activity)
Solubility:Not completely soluble in water, 2-10% insoluble material. Soluble portion of 1g dissolves in 3ml water; insoluble in usual organic solvents.
Ficin is a purified ficin preparation which is extracted from the latex of the fig tree Ficus glabrata. Ficin is classified as a thiol protease. Ficin hydrolyses the peptide bonds where the carbonyl group is from phenylalanine or tyrosine. When used in conjunction with other plants proteases, papain or bromelain, a synergistic effect may be observed.Immobilized Ficin was specifically designed for cleavage of mouse IgG1 into F(ab')2 or Fab fragments.The immobilization of ficin enhances stability against denaturation, heat and autolysis. Immobilization also eliminates any potential for antibody-enzyme adducts that cause continued sample digestion.
Ficin can be used to generate high yielding F (ab′)2 fragments from mouse IgG1. Ficin is classified as a thiol protease. It contains a single reactive cysteine at its active site. The amino acid homology of the active site is similar to that of papain. Ficin will cleave proteins at the carboxyl side of Gly, Ser, Thr, Met, Lys, Arg, Tyr, Ala, Asn, and Val. The reported Km for the chromogenic substrate pGlu-Phe-Leu-p-nitroanilide is 0.43 mM. Ficin is inhibited by iodoacetamide, iodoacetic acid, N-ethylmaleimide, mercuric chloride, DFP (diisopropyl fluorophosphate), TLCK (Na-p-Tosyl-lysine chloromethyl ketone), and TPCK (N-Tosyl-L-phenylalanine chloromethyl ketone).
