TA Instruments Nano ITC protocol Ian Kleckner Foster Lab
Contents
1. on e RPM 250 500 e Syringe select 100 or 250 depending on what was used e Setup Interval 300 sec 5 uL injection size e Auto equilibrate small medium or large heat expected default medium e Start e User manual is available on PC e Turn stirring OFF before removing syringe e Avoid vibration of instrument and table this will introduce noise into ITC signal e Do not get electronics wet e Do not bend the syringe e Do not drop the buret Printed 11 03 10 at 01 49 35 PM2. TA Instruments Nano ITC protocol lan Kleckner Foster Lab Equipment e Sample for cell 1 2 mL required for 0 950 mL active volume e Sample for syringe 300 or 750 uL for 100 or 250 uL desired active volume o Note there are two syringe sizes available 100 and 250 uL depending on how much ligand you wish you inject e Buffer 10 mL to rinse the cell before loading e 2 mini stir bars for degassing both samples Degassing e Must actively turn ON each of 4 steps at the degassing instrument e Set temperature used in experiment e Set stirring as fast as possible without creating vortex e Set vacuum to 600 mmHg maximum 635 mmHg e Set timer to 15 min e Caution it is possible to evaporate volatile solvent hence limit to 15 min Cleaning e Rinse with H20 is typically enough e Can also wash with 5 Contrad 70 detergent of 5 10 formic acid e Let this sit in the cell for 20 30 min and then rinse with H2O e Cleaning apparatus o WATER IN to H20 source o WATER OUT to waste bucket o VACUUM to back of degassing station Menu vacuum clean to turn on ae same a n Lega Q fie oan a 7 F pane g yi soak LTC re La C WN Software ITC Run e Check green light ITC is on e To check if experiment is running look for Injection at top right of screen e To stop experiment o Click STOP button o I Uncheck Stirrer on o Buret move up Loading cell e Rinse cell with buffer 3
3. times e Load cell with sample e It can overflow the cup before inserting the syringe unlike the VP ITC but this is unnecessary Printed 11 03 10 at 01 49 35 PM Reference cell e Fill with degassed water 1 0 mL do not overfill e The plug and dummy needle can be removed inserted with tweezers Loading syringe ligand e Wash with H20 and or alcohol to help dry e Air dry can use canned air if it is clean e Load sample into syringe Draw sample from source using the syringe plunger Remove plunger and allow sample to drain to back of syringe Cap syringe with plunger to trap 2 5 uL of air between plunger tip and sample Push plunger to expel air in the syringe Draw sample from source to fill remaining syringe volume plunger at absolute end of syringe Screw into buret and ensure that some sample is driven out by the buret Not shown wipe off excess volume from the tip of syringe with KimWipe 1 Vics aca MA a T YEr aie anya US h i Y 6 s N screw Ibo ail f AY bure ads a eme T the Seale Ol e a Da AOD egel afr Puna a ania if needle Gait suri Ae J SF Te Ql SOY VIAY Sarit e vanger ps G5 iqvid is at absolue ed FC MUSH COVQ Tout 420 Insert buret into cell e Insert this into the base in a vertical fashion do not bend needle e Push down and turn clockwise to engage spring loaded lock Software to set up run ITC Run e Check online green circle e Check that stirrer is
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