| User Guide for Reversed-Phase ZipTip Pipette Tips - Packard MutiPROBE Compatible | User Guide - P36404- > Product Catalogue | |
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Introduction
The ZipTip ® pipette tip is a 10 µL pipette tip with a bed of chromatography media fixed at its end. It is ideal for concentrating and puriying peptides, proteins or oligo-nucleotides prior to mass spectrometry,HPLC, capillary electrophoresis, and other analytical techniques.
Millipore, in partnership with Packard BioSciences, has developed a ZipTip configuration specific for the MultiPROBE II Liquid Handling Systems. The pipette tip contains a micro bed of C18 reversed-phase resin for peptide purification and concentration.
NOTE: For specific automation protocols and technical notes, visit the Millipore automation web site: www.millipore.com/automation.
Materials
The following table outlines the solutions required for use with ZipTip µ-C18 pipette tips.
| Solution | Description |
| Wetting solution | 50% acetonitrile (ACN) in Milli-Q ® grade water |
| Sample preparation solution | 0.1% trifluoroacetic acid (TFA) in Milli-Q ® grade water; final sample solution pH<4 |
| Equilibration solution | 0.1% TFA in Milli-Q ® grade water |
| Wash solution | 0.1% TFA in Milli-Q ® grade water |
| Elution solution* | 0.1% TFA/50% ACN, with or without MALDI matrix |
Overview of ZipTip Protocol Steps
Packard Deck Layout and Operation Protocols
Visit the Millipore automation web site at www.millipore.com /automation for a detailed description of the Packard BioScience MultiPROBE II system
deck layout along with instrument software protocols that can be downloaded.
Chemical Compatibility
. = Acceptable. Long exposures at room temperature have no significant effect.
? = Questionable. Short exposures at room temperature cause little or no damage.
X = Not recommended. Short exposure may cause permanent damage.
| Reagent | ZipTip |
| Acetic Acid (Glacial) | 4 |
| Acetone | x |
| Acetonitrile (100%) | 4 |
| Aliphatic Esters | ? |
| Ammonium Hydroxide (5% | 4 |
| Benzene (100%) | x |
| n-Butanol (100%) | 4 |
| Butyl Acetate | x |
| Chloroform (1%) | ? |
| Dichlorobenzene (100%) | x |
| Dichloromethane (1%) | ? |
| Diethanolamine (5%) | 4 |
| Dimethyl Acetamide (100%) | x |
| Dimethyl Formamide (100%) | x |
| Dimethyl Formamide (1%) | 4 |
| Ethanol (100%) | 4 |
| Formic Acid (5%) | 4 |
| Guanidine HCl (6 M) | 4 |
| Hydrochloric Acid | 4 |
| Hydrogen Peroxide | 4 |
| Isopropyl Alcohol (100%) | 4 |
| Mercaptoethanol (0.1 M) | 4 |
| Mercaptoethanol (1.0 M) | ? |
| Methyl Alcohol (100%) | 4 |
| Methyl Ethyl Ketone (100%) | x |
| Methyl Isobutyl Ketone (100%) | 4 |
| Nitric Acid (0.1 N) | 4 |
| Nitric Acid (1.5 N) | 4 |
| Phenol (0.5%) | x |
| Phosphoric Acid (1M) | 4 |
| Sodium Azide (1%) | 4 |
| Sodium Hydroxide (0.5 N) | 4 |
| Sodium Hydroxide (0.1 N) | 4 |
| Sodium Hypochlorite (100 ppm) | 4 |
| Sodium Hypochlorite (200 ppm) | 4 |
| Sulfuric Acid (1%) | 4 |
| Toluene (1%) | x |
| Triton® X-100 | 4 |
| Tween® | 4 |
| Urea (6 M) | 4 |
Troubleshooting
Sample preparation problems with ZipTip pipette tips can be divided into two categories:
- sample does not bind
- sample binds but is not recovered
problems. The procedures may or may not work with your ZipTip application.
For additional troubleshooting information and answers to Frequently Asked Questions, visit the Millipore Automation web site at www.millipore.com/automation.
Incomplete Binding
| Possible Cause | Suggested Procedure |
| C18 beads dewetted before sample was applied. The hydrophobic beads can de-wet in less than a minute. | After wetting with ACN, flush the with 0.1% TFA and leave the plug immersed in liquid until immediately before sample binding. |
| Sample was not sufficiently acidified with TFA. The pH should be below 4. The TFAconcentration should bebetween 0.1–1.0%. | Spike sample with a few microliters of 0.5–1% TFA. |
Incomplete Elution
| Possible Cause | Suggested Procedure |
| Sample tenaciously adsorbed to the C 18 particles. | Increase acetonitrile content of desorption solution to a maximum 75–90% ACN (v/v) in 0.1% TFA. |
| Sample not freely soluble in ACN. | Decrease ACN concentration to 20–40% in a 0.1% TFA or suitable ion-pairing agent. |
Specifications
Materials of Construction
Pipette tip: Polypropylene
Media: C 18 : spherical silica, 15 µm, 200Å pore size
Tip volume capacity 25 µL
Adsorptive bed 0.2 µL
Length 46 mm (1.825 in.)
Capacity (when used with saturating amounts of analyte): typically 2.0 µg
Max. Temperature 70 °C
Min. Temperature 4 °C
Working pH Range 2 to 13
Ordering Information
| ZipTip Pack | Resin Type | Catalogue Number |
| 96 pack, tip rack | C 18 (micro bed) | ZTC1 8P0 96 |
For a complete listing of available ZipTip chemistries, visit www.millipore.com/ziptip or contact your local Millipore office.
Technical Assistance
For more information, contact the Millipore office near-est you. In the U.S., call 1-800-MILLIPORE (1-800-645-5476). Outside the U.S., look in the Milli-pore
laboratory catalogue for the phone number of the Millipore office nearest you. You can also e-mail us at tech_service@millipore.com or visit our website (www.millipore.com).
Millipore’s Standard Warranty
Millipore Corporation (“Millipore”) warrants its products will meet their applicable published specifications when used in accordance with their applicable instructions for a period of one year from shipment of the products. MILLIPORE MAKES NO OTHER WARRANTY, EXPRESSED OR IMPLIED. THERE IS NO WARRANTY OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE. The warranty provided herein and the data, specifications and descriptions of Millipore products appearing in Millipore’s published catalogues and product literature may not be altered except by express written agreement signed by an officer of Millipore. Representations, oral or written, which are inconsistent with this warranty or such publications are not authorized and if given, should not be relied upon.
In the event of a breach of the foregoing warranty, Millipore’s sole obligation shall be to repair or replace, at its option, the applicable product or part thereof, provided the customer notifies Millipore promptly of any such breach. If after exercising reasonable efforts, Millipore is unable to repair or replace the product or part, then Millipore shall refund to the customer all monies paid for such applicable product or part. MILLIPORE SHALL NOT BE LIABLE FOR CONSEQUENTIAL, INCIDENTAL, SPECIAL OR ANY OTHER DAMAGES RESULTING FROM ECONOMIC LOSS OR PROPERTY DAMAGE SUSTAINED BY ANY CUSTOMER FROM THE USE OF ITS PRODUCTS.