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.
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
William R
WR
Kenealy
Thomas W
TW
Jeffries
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