Rapid 2,2'-bicinchoninic-based xylanase assay compatible with high throughput screening.
High-throughput screening requires simple assays that give reliable quantitative results. A microplate assay was developed for reducing sugar analysis that uses a 2,2'-bicinchoninic-based protein reagent. Endo-1,4-beta-D-xylanase activity against oat spelt xylan was detected at activities of 0.002 to 0.011 IU ml(-1). The assay is linear for sugar concentrations from 0 to 86 microg ml(-1) and can also be used to assay protein concentrations (0 to 143 microg ml(-1)) on the same plate. A variety of temperatures and pH conditions can be used and, after incubation, the assay requires only one detection reagent and one heating step.
Kenealy
William R
WR
Institute for Microbial and Biochemical Technology, USDA Forest Service, Forest Products Laboratory, One Gifford Pinchot Drive, Madison, WI 53726-2398, USA. bkenealy@fs.fed.us
Jeffries
Thomas W
TW
D002241:Carbohydrates / Q000032:analysis; D043364:Endo-1,4-beta Xylanases / Q000737:chemistry*; D004789:Enzyme Activation; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D025941:Protein Interaction Mapping / Q000379:methods*; D011506:Proteins / Q000032:analysis; D011804:Quinolines; D012710:Serum Albumin, Bovine / Q000032:analysis*; D014990:Xylans / Q000737:chemistry*; D014994:Xylose / Q000032:analysis*
D002241:Carbohydrates; D011506:Proteins; D011804:Quinolines; D014990:Xylans; C024795:bicinchoninate reagent; D012710:Serum Albumin, Bovine; D014994:Xylose; D043364:Endo-1,4-beta Xylanases