User:Daniel-Mario Larco/Notebook/AU Biodesign Lab - 09/03/2013/2014/04/15

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Objective Test HRP pellet Spin myoglobin and HB to see if it breaks apart UV/Vis myoglobin and Hb, spin, test again Procedure Make 0.1g/mol solutions of all three proteins in 50mM Tris buffer Spin solutions at 19000 rpm for 3 hours Test for HRP activity in the pellets Make OPD solution (1mg/mL) Make H2O2 solution 113uL 30% H2O2 in 887uL H2O Make mixture to be tested 2.025mL OPD solution 0.75mL H2O2 solution Take UV-Vis Add 0.225mL Data UV-Vis of Hemoglobin solution before Spin UV-Vis of Myoglobin solution before Spin UV-Vis of AuNP solution before spin Used for concentration calculations and size The last three images represent the activity of the pellets of HRP:AuNP conjugates for the three methods Notes When the tubes were removed from the centrifuge, none of the solutions had separated, pelleted out. There was a homogenous mixture for all three. The tubes were stored in the fridge over night to test for activity.