I27O is a 94kD polyprotein composed of eight repeats of the I27 domain of human fibronectin. It is used for the calibration of and as a reference for force extension experiments using atomic force microscopes. It is supplied in a ready-to-use solution. The protein has durable elasticity that allows for repeated unfolding and refolding. Single molecule measurements give up to eight saw-tooth force-extension curves (see Fig. 2) with the specifications below: Distance between peaks: 24.1 0.34nm Force peaks: 204 26pN Persistence length: 0.30 0.07nm Contour length: 24.1 0.34nm Instructions for Use: 1. Dilute the stock protein solution to a concentration of 100ug/ml in PBS pH 7.3 buffer and apply the sample to freshly evaporated gold coverslips. 2. Attach coverslips to the piezoelectric positioner stage. ] Note: The I27O was engineered with an amino-terminal hex-His peptide sequence and carboxy-terminal di-Cys. This configuration is hypothesized to allow the protein molecules to attach to the coverslip at the carboxy-terminal leaving the amino-terminus free to attach to the silicon nitride cantilever tip, when the tip is brought in close proximity to the absorbed protein. It is this model which is believed to account for the high frequency (>50%) of observing all eight domain extensions after each retraction as well as the ability to perform extension and retraction experiments on a single molecule. Storage and Stability: Aliquot to avoid repeated freezing and thawing and store at -70C. Aliquots are stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Purity:
Highly purified
Form:
Supplied as a liquid in 50mM Na2HPO4, 300mM NaCl, 20mM imidazole, pH 8.0.
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