The silicification reactions were performed according to our
prior methods [1,21]. The lyophilized 6mer and chimera proteins
were dissolved in double distilled water and then filtered with
5.0 lm syringe filters (Millipore, MA) to remove insoluble precipitates.
The protein concentration was measured by bicinchoninic
acid assay (Thermo Scientific, MA). 174 ll of 0.5 mg ml–1 protein
solutions was buffered to pH 7.0 by adding 13 ll of 1 M bis-tris
propane and 7 ll of 1 M citric acid buffers. 6 ll of prehydrolyzed
tetraethoxysilane (TEOS) solution (prepared by mixing 2.23 ml of
TEOS (Sigma-Aldrich, MO) with 7.76 ml of 50% ethanol/water solution,
100 ll of 1 M HCl and standing at room temperature for
15 min) was added and mixed thoroughly. The condensed and
aggregated silk–silica precipitates were collected after 1 h by centrifugation
at 13,000 rpm for 5 min and washed twice with water
and finally freeze-dried.
The silicification reactions were performed according to ourprior methods [1,21]. The lyophilized 6mer and chimera proteinswere dissolved in double distilled water and then filtered with5.0 lm syringe filters (Millipore, MA) to remove insoluble precipitates.The protein concentration was measured by bicinchoninicacid assay (Thermo Scientific, MA). 174 ll of 0.5 mg ml–1 proteinsolutions was buffered to pH 7.0 by adding 13 ll of 1 M bis-trispropane and 7 ll of 1 M citric acid buffers. 6 ll of prehydrolyzedtetraethoxysilane (TEOS) solution (prepared by mixing 2.23 ml ofTEOS (Sigma-Aldrich, MO) with 7.76 ml of 50% ethanol/water solution,100 ll of 1 M HCl and standing at room temperature for15 min) was added and mixed thoroughly. The condensed andaggregated silk–silica precipitates were collected after 1 h by centrifugationat 13,000 rpm for 5 min and washed twice with waterand finally freeze-dried.
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