Ficin - Sigma-Aldrich
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
FICIN ACTIVITY DETERMINATION - Animal Science
FICIN ACTIVITY DETERMINATION Updated September 2013 I Personal Protective Equipment: A Lab coat B Safety glasses/goggles C Latex gloves II Reagents: A Denatured hemoglobin powder 1 Add 20 g denatured hemoglobin/liter assay buffer B Stock phosphate buffer, pH 6 0 5 M 1 Add 10 54 g Potassium Phosphate (K 2 HPO 4) (dibasic
Fab & F(ab) Fragmentation of Mouse IgG
Ficin is typically used to cleave mouse IgG 1 as this are difficult to cleave with papain and pepsin In the presence of 1mM or 10mM cysteine, ficin generates F(ab;) 2 and Fab fragments respectively Im mobilized Ficin is a convenient reagent for producing Fab and F(ab ;) 2 fragments as it avoids the need to remove the ficin enzyme after digestion
Blood Groups Duffy, and MNSs Group Systems
Duffy antigens can be destroyed by enzymes such as ficin, papain, bromelain, chymotrypsin, ZZAP When compared to Rh or Kell antigens, Duffy antigens are not very immunogenic So, anti-Fya or anti-Fyb is not common Fy (a-b-) is not Fy null, but homozygous for Fyb gene, they express Fyb antigen in other tissues, but not on RBCs → only
Blood Groups Kell Group - UCLA Health
papain, ficin DTT and ZZAP denature all Kell antigens, but not Kx Immunogenicity: strongly immunogenic - One unit of K+ blood transfused to K- patients -70 of patients will have anti-Kell (in comparison to 80 for D- patients receiving D+ blood)
Roche Applied Science The Complete Guide for Protease Inhibition
(e g , Bromelain, Ficin, Papain) Trypsin Protease-specific for the inhibitors inhibition of: Protease Inhibitors from Roche Applied Science —
HTLA Antibodies: Identification and Management Strategies and
Ficin and DTT red cells -Antibody is to an antigen destroyed by ficin, but not by DTT -Titers to a dilution of 32 -Reacts similar with PEG, LISS, or without
Please Don’t Say HTLA
Ficin destroys Ch, Rg, JMH Trypsin & DTT destroy Knops & Dombrock Other Useful Techniques Antibody neutralization: Plasma inhibition for Ch/Rg
Complete - Immucor
Ficin-treated panel Includes one vial each of Diluent, Ficin Solution and : Ficin Control Ficin enhances certain antibodies and destroys certain antigens to aid :
Antibody Identification Case Studies
Test same panel ficin‐treated (Some alloantibodies have already been excluded with cell #3) DC E c eLea Leb P 1 MN S s K kFya Fyb Jka Jkb PEG IAT Ficin IAT 1+ +0 0+0+ + + 0+0 0 +0+0+ 3+ 2+ 2+ +0 0+0+ +0+ + +0+ + 0 0 + 3+ 2+ 30 +0+ + 0+ +0+0+0+0+ + 0 0 0 4+ 0 + +0 0 0 +0+ + +0+0+0 + 3+ 2+ 5+ 0 0 +++ 0 +++++ 0 0 + 0 3+ 2+ 6+ +0+ + 0+0 + +0 +0+0
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