Agilent Nanoflow Proteomics Solution Quick
Contents
1. D C PFS 25 350 0 172 6 G1375 87322 15 Version Ill IV D C PFS 25 550 0 270 9 G1375 87323 16 Version IV C D PFS 25 100 0 049 2 G1375 87320 16 Version II Ill C D PFS 25 350 0 172 6 G1375 87322 16 Version II Ill C D PFS 25 550 0 270 9 G1375 87323 16 Version II Ill C D PFS 25 700 0 344 12 G1375 87324 Fitting A 5062 2418 Fitting B 5063 6593 5065 4423 Fitting C 5065 4410 Fitting D 5065 4422 G2228 90000 Agilent Technologies Inc 2002 Agilent Technologies Deutschland GmbH Hewlett Packard Strasse 8 76337 Waldbronn Germany Part Number G2228 90000 Edition 07 02 Printed in Germany2. eno Agilent Nanoflow Proteomics 0 Q o Solution 909 Quick Reference Guide r 8 The diagram below provides an overview of all capillaries needed for complete setup of an Agilent Nanoflow Proteomics Solution You can use any Agilent 1100 series pump as second pump For details on each of the capillaries see Table 1 on page 4 Second Pump Isocratic pump Quaternary pump E Binary pump E E aS Capillary pump a Analytical column lon trap TCC Thermostatted Column Compartement A Agilent Technologies Tips for Successful Operation of Nanoflow Proteomics Solution System For direct injection place the well plate sampler close to the lon Trap Note system pressure of your new installed system under typical conditions 40 50 bar at 300 nl min of water with a 50x0 075 mm 3 5 um column Capillaries Wash both ends with organic solvent and flush before connecting new capillaries to other components Always install or retighten without flow Use pH lower than 9 Compare capillary pressure drop according to table 1 replace it if you have more than 30 deviation Pump Degasser Use clean solvent bottles and solvent Use primary flow rate for low solvent consumption After changing solvents purge each channel for 4 min After sitting idle set composition to 50 ACN and pump until pressure ripple is less than 3 Check pressure drop of solvent filter in front of
3. t of a few hundred nA e Gently push the needle through the e Do not use highly conductive sample conductive ferrule until it touches the filter solvents e g 1 TFA to avoid arcs screen of the column and pull it back a needle damage little bit before handtighten e Do not leave the needle close to the plate e New needles sometimes need some without flow e g after a sequence use a positioning back and forwards to the plate method with low dry temperature until a good spray is obtained For more information on your Agilent Nanoflow Proteomics Solution please check the Nanoflow Proteomics Solution Getting Started Guide G4000 90020 and the Nano Pump User Manual G2226 90000 Nanoflow Proteomics Solution Quick Reference Guide Table1 Capillaries of the Agilent Nanoflow Proteomics Solution Item Fitting Material Diameter Length Volume P drop bar Part number type yim mm pl for 1 pl min H20 1 A A SST G1311 67304 2 A A SST G1312 67300 3 A A SST G1312 67302 4 A A SST G1312 67304 5 A A SST 250 130 6 381 01090 87308 6 B C PFS 75 650 2 872 0 3 G1375 87327 6a direct injection B C PFS 50 150 0 295 G1375 87300 7 C 2000 G1375 87326 8 C B PFS 100 200 1 570 G1375 87312 9 B D PFS 100 1100 8 639 G1375 87315 10 Needle SST G1375 87201 11 Needle seat G1375 87101 12 C D PFS 25 100 0 049 2 G1375 87320 12 C D PFS 50 100 0 196 G1375 87325 13 A A SST 170 280 6 355 G1375 87400 14 D D PFS 25 220 0 108 4 G1375 87321 15 Version I Il
4. the EMPV once a month After sitting idle for a day or longer flush each channel for a few minutes Check for plugged column capillaries if pressure increases more than 30 Allow enough trapping injection time for the sample to be transferred to the trapping or analytical column Avoid gaps within fittings Do not overtighten trap in module doors or bend with radius smaller than 4 cm Inspect suspicious capillary milky surface under microscope Replace capillaries with permanent sharp bends System backpressure should be higher than 20 bar Irregular flow pressure fluctuations indicate partially blocked capillaries Regular fluctuations indicate air within the high pressure path Rotate EMPV valve once while under flow to remove dirt from the valve seat Fast composition change is not used for Nanoflow proteomics solution Never run without solvent inlet filters Use glass bottled solvents Nanoflow Proteomics Solution Quick Reference Guide Well plate sampler WPS e Cool sample e Ensure comparable pressure drop ina e Use needle wash mainpass and bypass once a week e Prime and verify wash pump once a week e Use bottom sensing for minimum sample e Check alignment once a month solvent e For direct injection use bypass mode allowing 3 6 min 300 nl min sample transfer between WPS and column lon Trap e Cut and rinse sprayer needle before e Set voltage to about 1400 V to generate a installation curren
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