Thermo Scientific Aspire RP30 Desalting Tips
Contents
1. manufacturer or third party supplier to the extent such assignment is allowed by such original manufacturer or third party supplier In no event shall Seller have any obligation to make repairs replacements or corrections required in whole or in part as the result as i normal wear and tear ii accident disaster or event of force majeure ii1 misuse fault or negligence of or by Buyer iv use of the Products in a manner for which they were not designed v causes external to the Products such as but not limited to power failure or electrical power surges vi improper storage and handling of the Products or vii use of the Products in combination with equipment or software not supplied by Seller If Seller determines that Products for which Buyer has requested warranty services are not covered by the warranty hereunder Buyer shall pay or reimburse Seller for all costs of investigating and responding to such request at Seller s then prevailing time and materials rates If Seller provides repair services or replacement parts that are not covered by this warranty Buyer shall pay Seller therefore at Seller s then prevailing time and materials rates Any installation maintenance repair service relocations or alteration to or of or other tampering with the products preformed by any person or entity other than seller without sellers s prior written approval or any use of replacement parts not supplied by seller shall immediately void and can2. of Equilibration Wash Solution and expel solution back into the same tube Repeat once with same fraction of Equilibration Wash Solution row B Keeping the speed and STEPPER mode adjust pipette volume setting equal to the sample volume and insert Aspire tip s into the yellow tube row C Bind peptides by aspirating and dispensing diluted sample at least 6 times Expel flow through back into the same tube Note Retain flow through fraction if verification of sample binding is desired Keeping the speed and STEPPER mode change pipette to the volume setting to 100 ul Insert Aspire tip s into the second green tube row D Wash resin by aspirating 100 ul of Equilibration Wash solution Expel solution to same tube Repeat once with the same fraction of Equilibration Wash Solution row D Insert Aspire tip s into the third green tube row E Repeat wash step outlined in step 10 Keeping the speed and STEPPER mode adjust pipette volume setting equal to the elution volume Insert Aspire tip s into the orange tube row F Elute peptides by aspirating and dispensing Elution Solution 5 times Dispense eluate back into the same tube Note Although bubbling may occur during these cycles it will not affect sample recovery Continue elution by inserting Aspire tip s into the orange tube row G Aspirate and dispense Elution Solution 5 times Dispense eluate into the first elution fraction in order to pool the two fractions together The ora
3. the provided rack as illustrated in Diagram 1 Each sample requires the following quantities of color coded tubes for processing e 1 blue 3 green e 1 yellow e 2 orange Note Up to 12 samples can be processed in parallel on a multichannel pipette OC MOOK Activation Solution Equilibration Wash Solution Diluted Peptide Sample Elution Solution Sample 1 Sample 2 Diagram 1 Tube arrangement for processing of twelve samples 10 11 I 13 14 15 Aliquot 100 ul of Activation Solution into each blue tube Aliquot 100 ul of Equilibration Wash Solution into each green tube In each yellow tube mix 3 parts sample to 1 part Sample Solution The final sample will contain 0 5 TFA in 5 ACN For example Add 25 ul of Sample Solution to 75 ul of sample Aliquot a minimum of 25 ul of Elution Solution into each orange tube The recommended range of elution volumes is 25 50 ul Adjust pipette setting to 100 ul and attach Aspire Tip s to the pipette Completely re hydrate and activate resin inside the Aspire tip s by aspirating 100 ul of Activation Solution from the blue tube row A Expel solution back into the same tube Repeat once with the same fraction of Activation Solution row A Insert the Aspire tip s into the green tube row B Equilibrate resin by aspirating 100 ul of Equilibration Wash Solution and expel solution back into the same tube Repeat once with same fraction of Equilibration
4. 3 U K and 4 Australia MSDS Information The Material Safety Data Sheet MSDS information for the products is provided on www thermo com Simply navigate to the product page to retrieve any associated MSDS s in a print format MSDS documents are not included with product shipments Expiration Period One year from date of sale for product used handled and stored according to Manufacturer instructions see details under Warranty The most current versions of all Aspire product instructions are available at www thermo com For a copy please go to www thermo com or your local distributor Thermo Fisher Scientific Inc 12 2008 All rights reserved All trademarks are the property of Thermo Fisher Scientific Inc and its subsidiaries Specifications terms and pricing are subject to change Not all products are available in all countries Please consult your local sales representative for details Printed in the USA Warranty Sellers warrants that the Products will operate or perform substantially in conformance with Seller s published specifications and be free from defects in material and workmanship when subjected to normal proper and intended usage by properly trained personnel for the period of time set forth in the product documentation published specifications or page insert If a period of time is not specified in Seller s product documentation published specifications or package inserts the warranty period shall be one 1
5. 5X Repeat with elution fraction two Combine elution fractions by finally dispensing fraction two into the same tube used to collect fraction one Novus electronic pipette Adjust the pipette volume setting equal to the volume of the elution fractions t DRY and RESUSPEND for MS ANALYSIS Piace entire arange elution collection tube inside of a 1 5 ml ar 2 mi microcentrifuge tube Proceed with drying in a vacuum evaporator put A iil il A i iil mi m sil ili l ni wi inl aaarnas l aa vl nv pishe MN i ji LS Troubleshooting High pH lack of ion pairing Ensure sample is diluted in Poor or agents Sample Solution with TFA incomplete sample Sample contains organic Dry sample and resuspend in binding solvent 20 ul 0 1 0 5 TFA Peptides are too hydrophilic Increase TFA concentration to bind resin from 0 5 to 1 in the sample binding step Highly hydrophobic sample Use 70 95 ACN 0 1 Poor or TFA for elution conditions incomplete sample Sample contains Increase TFA concentration recovery hydrophilic peptides or a from 0 5 to 1 in the wide range of peptides sample binding step Related Thermo Scientific Products 4630040X Thermo Scientific Finnpipette Novus Multichannel Electronic Pipette 8 channel 30 300 ul 4630050X Thermo Scientific Finnpipette Novus Multichannel Electronic Pipette 12 channel 30 300 ul X 0 Eur 1 U S 2 Japan
6. Thermo Scientific Aspire RP30 Desalting Tips Catalog number Description 990 01 Thermo Scientific Aspire RP30 Desalting Tips 96 purifications Introduction The Thermo Scientific Aspire RP30 Desalting Tips purify desalt and concentrate peptides or low molecular weight proteins 65 kDa or less The Aspire Tip is a 200 ul chromatography pipette tip which contains a bed of proprietary dehydrated porous Styrene divinylbenzene SDVB reversed phase resin This SDVB resin offers superior overall peptide binding and recovery compared to traditional C18 products Mass spectrometry methods are becoming important tools for the study of biological compounds Salts detergents and other contaminants present in biological samples have deleterious effects on the quality of MS spectra Chromatography tips are convenient for peptide desalting and have been widely used for Matrix assisted laser desorption ionization MALDI sample cleanup and concentration As researchers begin to analyze samples of increasing complexity sample cleanup using conventional desalting tips becomes less ideal due to their low binding capacity 1 5 ug and recovery The Aspire RP30 Desalting Tips allow purification of biological samples including complex mixtures containing up to 30 ug of total peptides rendering them ideal for pre LC MS sample cleanup The Aspire Tips effectively remove salts detergents and other ion suppressing interference from biological samples prior to LC MS a
7. Wash Solution row B Adjust pipette setting to sample volume and insert Aspire tip s into the yellow tube row C Bind peptides by aspirating and dispensing diluted sample at least 6 times Expel flow through back into the same tube Note Retain flow through fraction if verification of sample binding is desired Adjust pipette setting to 100 ul Insert Aspire tip s into the second green tube row D Wash resin by aspirating 100 ul of Equilibration Wash solution Expel solution to same tube Repeat once with same fraction of Equilibration Wash Solution row D Insert Aspire tip s into the third green tube row E Repeat wash step outlined in step 10 Adjust pipette setting to 5 ul greater than the elution fraction volume For example Adjust pipette setting to 30 ul for the elution fraction volume of 25 ul Insert Aspire tip s into the orange tube row F Elute peptides by aspirating and dispensing Elution Solution 5 times Dispense eluate back into the same tube Note Although bubbling may occur during these cycles it will not affect sample recovery Continue elution by inserting Aspire tip s into the orange tube row G Aspirate and dispense Elution Solution 5 times Dispense eluate back into the first orange tube in row F to pool the two elution fractions The orange tubes can be securely sealed with the provided caps If needed gently dry sample in a vacuum evaporator to further concentrate eluate and lower the acetonitri
8. cel all warranties with respect to the affected products The obligations created by this warranty statement to repair or replace a defective product shall be the sole remedy of buyer in the event of defective product except as expressly provided in this warranty statement seller disclaims all other warranties whether express or implied oral or written with respect to the products including without limitations all implied warranties of merchantability or fitness for any particular purpose Seller does not warrant that the products are error free or will accomplish any particular result
9. le concentration for downstream applications To dry eluate carefully insert the orange tube pooled elution fraction without the cap into a 1 5 or 2 0 ml centrifuge tube and proceed drying in a vacuum evaporator For LC MS applications resuspend sample in 0 1 formic acid or the appropriate solution after drying For MALDI MS analysis carefully resuspend sample in 1 2 ul of matrix solution prepared just before use Procedure Using a Thermo Scientific Finnpipette Novus Electronic Multichannel Additional Materials Required 200 300 ul single or multichannel electronic Novus pipette Ultrapure water Acetonitrile ACN Trifluoroactic acid TFA Methanol e gt Material Preparation Activation Solution 50 Methanol Note ACN can be substituted for methanol Equilibration Wash Solution 0 5 TFA in 5 ACN Sample Solution 2 TFA in 20 ACN Elution Solution 70 ACN e o o o y C Purification Protocol Note Perform the following procedure at room temperature 1 Arrange the color coded tubes in the provided tube rack as illustrated in Diagram 1 page 2 Each sample requires the following quantities of color coded tubes for processing e 1 blue 3 green 1 yellow 2 orange Note Up to 12 samples can be processed in parallel on a multichannel pipette 2 With the Finnpipette Novus electronic multichannel pipette aliquot 100 ul of Activation Solution into each blue tube 3 Aliquot 100 ul of Equilib
10. nd LC MS MS analysis improving sensitivity and data quality The color coded multichannel purification protocol is optimized for low to medium throughput sample cleanup The Aspire tips are compatible with a variety of handheld pipettes For the user s convenience the procedure is further optimized on a Thermo Scientific Finnpipette Novus electronic multichannel pipette see page 4 for instructions Twelve samples can be processed in parallel within 20 minutes without the need for centrifugation Kit Contents Aspire RP30 Desalting Tips 96 tips Assorted color coded tubes Tube rack 1 Tube caps 96 User manual Storage Upon receipt store all components at room temperature Product is shipped at ambient temperature This product is supplied for life science research use only This product is not intended for medicinal diagnostic or therapeutic use e o o o yj C eo ie gt Procedure Using a Handheld Pipette Additional Materials Required 200 300 ul single or multichannel pipette manual or electronic Ultrapure water Acetonitrile ACN Trifluoroacetic acid TFA Methanol Material Preparation Activation Solution 50 Methanol Note ACN can be substituted for methanol Equilibration Wash Solution 0 5 TFA in 5 ACN Sample Solution 2 TFA in 20 ACN Elution Solution 70 ACN Purification Protocol Note Perform the following procedure at room temperature Arrange the color coded tubes in
11. nge tubes row F can be securely sealed with the provided caps If needed gently dry sample in a vacuum evaporator to further concentrate eluate and lower the acetonitrile concentration for downstream applications To dry eluate carefully insert the orange tube pooled elution fraction without the cap into a 1 5 or 2 0 ml centrifuge tube and proceed drying in a vacuum evaporator For LC MS applications resuspend sample in 0 1 formic acid or the appropriate solution after drying For MALDI MS analysis carefully resuspend sample in 1 2 ul of matrix solution prepared just prior to use D Procedure Flowchart PREPARE TUBE RACK Arrange the color coded tubes in the provided rack Fill tubes with the appropriate volumes of solutions t ACTIVATE Row A blue tubes Completely re hydrate the resin bed by aspirating and dispensing activation solution 2X Novus electronic pipette Use STEPPER mode and adjust the volume to 100 pl T EQUILIBRATE Row B green tubes Equilibrate the resan by aspirating and dispensing equilibration wash solution 2X f BIND Row C yellow tubes S EE PE DA E AAE electronic pipette Adjust the volume setting equal to volume of the sample fraction t WASH Row D and E green tubes Aspirate and dispense wash fraction one 2X with wash fraction two Novus electronic pipette Adjust the volume setting to 100 pl ELUTE Row F and G orange tubes Aspirate and dispense elution fraction one
12. ration Wash Solution into each green tube 4 Ineach yellow tube mix 3 parts sample to 1 part Sample Solution The final sample will contain 0 5 TFA in 5 ACN For example Add 25 ul of Sample Solution to 75 ul of sample 5 Aliquot a minimum of 25 ul of Elution Solution into each orange tube The recommended range of elution volumes per tube is from 25 50 ul 6 Using the STEPPER function on the Novus pipette adjust pipette setting to 100 ul Adjust the aspirate and dispense rates to 9 on the Novus pipette Attach Aspire RP30 Desalting Tip s to the pipette Speed Function Total Volume of Steps Figure 1 Finnpipette Novus electronic multichannel STEPPER function settings Note The STEPPER function is used as it allows for a blowout step which ensures complete dispensing of all aspirated liquid 7 10 11 12 13 14 15 16 Completely re hydrate and activate resin inside the Aspire tip by aspirating 100 ul of Activation Solution from the blue tube row A Expel solution back to the same tube Repeat once with same fraction of Activation Solution row A Note Hold the trigger button down during the blowout step until all of the liquid is expelled from the tip The dispensing step only expels a small portion of the liquid the blowout step subsequently expels the remainder of the liquid from the tip Insert Aspire tip s into the green tube row B Equilibrate resin by aspirating 100 ul
13. year from the date of shipment to Buyer for equipment and ninety 90 days for all other products the Warranty Period Seller agrees the Warranty Period to repair or replace at Seller s option defective Products so as to cause the same to operate in substantial conformance with said published specifications provided that Buyer shall a promptly notify Seller in writing upon the discovery of any defect which notice shall include the product model and serial number if applicable and details of the warranty claim and b after Seller s review Seller will provide Buyer with service data and or a Return Material Authorization RMA which may include biohazard decontamination procedures and other product specific handling instructions then if applicable Buyer may return the effective Products to Seller with all costs prepaid by Buyer Replacement parts may be new or refurbished at the election of Seller All replaced parts shall become the property of Seller Shipment to Buyer repaired or replacement Products shall be made in accordance with the Delivery provisions of the Seller s Term and Conditions of Sale Consumables are expressly excluded this warranty Notwithstanding the foregoing Products supplied by Seller that are obtained by Seller from an original manufacturer or third party supplier are not warranted by Seller but Seller agrees to assign to Buyer any warranty rights in such Product that Seller may have from the original
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