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Protein purification- A serine protease present in the venom was purified by liquid phase isoelectric focussing using a micro rotofor (Bio-Rad, Hemel Hempstead, UK) followed by Superdex 75 gel filtration using a 16 mm ×70 cm column.
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Samples were applied using a Micro dispenser (DRUMMOND).
The ear thickness was measured using a micro caliper (Mitutoyo, Kanagawa, Japan) 24 hr after injection.
The absorbance was measured at 492 nm using a micro plate reader (Model 680, Bio-Rad).
Peptide concentration was determined using a micro bicinchoninic acid assay (Micro BCA Assay; Thermo Scientific).
Micro-hardness was examined using a micro-hardness tester.
The robot's blinking eyes and mouth are controlled using a micro-controller and a PC.
Hybridization was performed using a micro-circulation pump (Atactic Technologies).
Protein concentration was estimated using a micro-BCA kit (Thermo Scientific, Rockford, IL) with a BSA standard.
Luminescence was read 15 minutes later using a Micro-beta (Perkin Elmer) plate reader.
ESCs were differentiated into osteoblast lineage using a micro-mass approach.
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