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proteins using AbCA agarose
A monoclonal antibody was developed that reacts only to the AviTag when
fused to the C-terminus of desired protein. It will identify C-terminus fusions
of AviTag on western blots ａnd has been immobilized on agarose beads for
purifications purposes (catalog #AbC agarose). The AviTag containing
protein can be eluted from the monoclonal antibody affinity column using
documented mild elution conditions which retaining protein activity Elution
buffer I(Thompson NE, Hager, DA ａnd Burgess, RR (1992) Biochemistry 31,
7003-7008; Thompson NE ａnd Burgess, RR (1994) Protein Expression and
Purification 5,468-475). If protein can withstand elution at pH 3.0, Elution
buffer II can be used.
Reagents provided by Avidity, LLC
Affinity purified IgG against AviTag fused to C-terminus of disired protein
crosslinked to agarose 10 mg IgG/ml resin in PBS containing 0.05%
sodium azide
20 ml column for your use. Comes with upper bed support that prevents
column from drying out, column plug ａnd cap.
Solutions: to be provided by user
PBS: 138 mM NaCl, 2.6 mM KCl in 10 mM potassium phosphate (pH 7.4)
PBST: PBS plus 0.05% Tween 20
Elution buffer I: 40% 1,2, propanediol, 0.75M Sodium pyruvate, 50 mM
Tris-HCl, 1 mM EDTA pH 7.5
Elution buffer II: 0.1 M glycine pH 3.0
