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文献和实验Mapping Protein Distributions on Polytene Chromosomes by Immunostaining
solution 2 can be raised to 500 mM. Problem: HRP staining is weak. [Step 30] Solution: Silver treatment of the preparation turns the brown color of a DAB signal to black, thus substantially improving the contrast
of the flowthrough was less than 0.01 (approximately 50 ml). Protein was eluted with a linear gradient of 15 ml each of 0 and 500 mM imidazole chloride in elution buffer (50 mM Na phosphate [pH 7.0], 100 mM NaCl, 10% glycerol). Fractions of 1 ml were collected
Thrombin Cleavage of GST-Fusion protein
activity keeps the enzyme in aliquots at -80°C. Thrombin (MW 37kDa) ~4500units/mg. One unit cleaves theoretical 100ug protein in 16hr at 22°C in buffer. Another possibility is to use high purity enzyme from Sigma # T-6884: 3-10 U of thrombin can be used
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