Bio-Rad ReadyPrep 2-D Cleanup Kit User Manual
ReadyPrep™2-D Cleanup Kit
Instruction Manual
Catalog #163-2130
For technical service, call your local Bio-Rad office, or
in the US, call 1-800-4BIORAD (1-800-424-6723)
Bio-Rad Laboratories, Inc.
2000 Alfred Nobel Dr.
Hercules, CA 94547 USA
(510) 741-1000
1-800-424-6723
4110143 Rev A
Table of Contents
Section 1 Introduction……………………………………...1
Section 2 Kit Specifications…………………………......2 –3
Section 3 Storage Conditions………………………..........3
Section 4 Instructions for Use………………….............4–9
Section 5 Appendix……………..……………............10–15
Section 6 Product Information………………….........16–17
Section 1
Introduction
The ReadyPrep 2-D cleanup kit facilitates the preparation
of low conductivity samples suitable for isoelectric focusing
(IEF) and 2-D gel electrophoresis. Additionally, the kit
concentrates proteins from samples that are too dilute,
allowing for higher protein loads that can improve spot
detection. The ReadyPrep 2-D cleanup kit can be used
with virtually any protein sample to generate improved 2-D
results by reducing streaking, background staining, and
other gel artifacts associated with substances
contaminating 2D/IEF samples. The entire process can be
completed in approximately one hour and does not
produce alterations in the isoelectric point of proteins. The
procedure works by quantitatively precipitating and
concentrating proteins in a sample while leaving behind
and washing away substances like ionic detergents, salts,
nucleic acids, lipids, and plant-derived phenolic
compounds, all of which are known to interfere with IEF.
The precipitation is not affected by the presence of
detergents, chaotropes or other common buffers, salts and
reagents used to extract or treat proteins. After
precipitation, the proteins are washed and then
resuspended in the IEF/2D-compatible rehydration/sample
buffer of choice, ready for isoelectric focusing or Laemmli
sample buffer, ready for SDS-PAGE.
1
Section 2
Kit Specifications
Each ReadyPrep 2-D cleanup kit provides sufficient reagent
to purify 50 samples of 100 µl, each sample containing up
to 500 µg of protein. The kit contains the components
listed below. Certificates of analysis or MSDS forms are
available upon request.
Items Supplied With Kit
One bottle containing 15 ml of precipitation agent 1*
One bottle containing 15 ml of precipitation agent 2
One tube containing 2 ml of wash reagent 1
One bottle containing 50 ml of wash reagent 2*
One tube containing 0.25 ml of wash 2 additive
One instruction manual
*Care should be exercised when handling precipitation
agent 1 and wash reagent 2.
Precipitation agent 1 is corrosive!
Wash reagent 2 is flammable and an irritant!
2
Items Required But Not Provided:
• 1.5 ml microcentrifuge tubes
• Microcentrifuge capable of spinning at 12-16,000 x g at
4°C.
• Rehydration/sample buffer (see Section 5 for recipes and
Section 6 for premixed, ready-to-use rehydration/sample
buffers)
• Vortex mixer
• ReadyPrep proteomic grade water or other ultrapure
water
• RC DC™ Protein Assay (Bio-Rad catalog #500-0121 or
#500-0122)
Section 3
Storage Conditions
Store the kit at room temperature. Wash reagent 2 must be
stored at -20°C for at least 1 hr prior to use. For
convenience, store wash reagent 2 in a -20°C freezer.
3
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