Thermo Fisher Scientific Pierce Top 12 Abundant Protein User Guide
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INSTRUCTIONS
Pierce™ Top 12 Abundant Protein
Depletion Spin Columns
85164 85165
Number Description
85164
2607.0
Pierce Top 12 Abundant Protein Depletion Spin Columns, 6 columns, each column contains
62% slurry in 10mM PBS, 0.15M NaCl, 0.02% sodium azide, pH 7.4
85165 Pierce Top 12 Abundant Protein Depletion Spin Columns, 24 columns, each column contains
62% slurry in 10mM PBS, 0.15M NaCl, 0.02% sodium azide, pH 7.4
Storage: Upon receipt store at 4
°
C. Product shipped with an ice pack. Do not freeze product.
Introduction
The Thermo Scientific Pierce Top 12 Abundant Protein Depletion Spin Columns are used for single-step removal of twelve high-abundance proteins from 10µL of human serum or plasma. The resin uses highly specific immobilized antibodies for protein removal, providing minimal nonspecific interactions with other proteins.
Plasma proteome studies are limited by the vast dynamic range (10 orders of magnitude) of protein abundance. Many proteins of pharmaceutical interest are present at low concentrations in the plasma and are difficult to detect in the presence of high-abundant proteins. The proteins listed in the table below constitute ~95% of all plasma proteins. The Pierce Top 12
Abundant Protein Depletion Spin Columns remove > 95% of these 12 high-abundant proteins from 10µL of serum or plasma samples and enable the identification and quantitation of low-abundant proteins in samples by mass spectrometry (MS).
Table 1. List of the top 12 proteins removed by the Thermo Scientific Pierce Top
12 Abundant Protein Depletion Spin Columns.
α1-Acid Glycoprotein
Fibrinogen
α1-Antitrypsin
Haptoglobin
α2-Macroglubulin
IgA
Albumin IgG
Apolipoprotein A-I IgM
Apolipoprotein A-II Transferrin
Important Product Information
•
The depletion spin columns contain a storage solution used as a dilution and binding buffer for direct processing of 10µL of serum or plasma.
Note: No other additions or solvent exchanges are required before protein depletion.
•
Each depletion spin column processes a maximum of 10µL of human serum or plasma. To process larger serum sample volumes, use multiple depletion spin columns. For serum samples containing abnormally high amounts of albumin or
IgG, the sample load may need to be reduced.
•
The depletion spin columns are designed for single use. Do not reuse the resin.
•
The depletion spin columns are designed for use with human serum or plasma samples and have not been tested on other species. Alternative human biological fluids (e.g., cerebrospinal fluid or amniotic fluid) may be used with this product, but may require optimization.
Pierce Biotechnology
3747 N. Meridian Road
PO Box 117
Rockford, lL 61105 USA
(815) 968-0747
(815) 968-7316 fax www.thermoscientific.com/pierce
Additional Materials Required
•
Microcentrifuge capable of operating at 1000 × g
•
2mL collection tubes
•
End-over-end mixer
Spin Column Procedure for Top 12 Protein Removal
Note: The concentration of serum/plasma proteins can vary widely depending on the origin of the sample. Each 810µL of antibody resin slurry is optimized to process 10µL (600µg) of serum or plasma. For best results, optimize the ratio of sample to slurry volume for each specific application.
1.
Equilibrate the depletion spin column to room temperature.
2.
Remove the column screw cap and add 10µL of sample directly to the resin slurry in the column.
3.
Cap the column and invert the column several times until the resin is completely suspended in the solution.
4.
Incubate the mixture in the column with gentle end-over-end mixing for 60 minutes at room temperature. Make sure the sample mixes with the resin during the incubation period. Alternatively, vortex every 5 minutes.
5.
Twist off the bottom closure and loosen the cap. Place column into a 2mL collection tube and centrifuge at 1000 × g for
2 minutes.
6.
Discard the column containing the resin.
7.
Filtrate contains sample with the top 12 proteins removed. Use for further processing or store for later use. The depleted sample will be in 10mM PBS, 0.15M NaCl, 0.02% azide, pH 7.4.
8.
Sample processing will depend on the type of downstream analysis and may require buffer exchange and/or concentration for 2D gel electrophoresis and MS analysis. Use Thermo Scientific Pierce Protein Concentrators for buffer exchanging and/or concentration (see Related Thermo Scientific Products Section).
Troubleshooting
Problem Possible Cause Solution
Incomplete removal of albumin or IgG
Sample exceeds binding capacity
Incomplete binding
Sample is not mixed during incubation
Reduce amount of sample processed
Increase incubation time
Mix the sample with resin by gentle end-over-end mixing and make sure that the sample is mixing with the resin during the incubation period.
Related Thermo Scientific Products
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85161
85162
31876
87748
23225
24590
84840
Pierce Top 2 Abundant Protein Depletion Spin Columns, 6 columns
Pierce Top 2 Abundant Protein Depletion Spin Columns, 24 columns
Pierce Normal Human Serum
Pierce Protein Concentrators, 9K MWCO, 7mL
BCA Protein Assay Kit
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Pierce MS Sample Prep Kit
General Reference
Anderson, N.L. and Anderson, N.G. (2002). The human plasma proteome: History, character, and diagnostic prospects. Mol Cell Proteomics 1: 845-67.
Pierce Biotechnology
3747 N. Meridian Road
PO Box 117
Rockford, lL 61105 USA
(815) 968-0747
(815) 968-7316 fax www.thermoscientific.com/pierce
2
Products are warranted to operate or perform substantially in conformance with published Product specifications in effect at the time of sale, as set forth in the Product documentation, specifications and/or accompanying package inserts (“Documentation”). No claim of suitability for use in applications regulated by FDA is made. The warranty provided herein is valid only when used by properly trained individuals. Unless otherwise stated in the Documentation, this warranty is limited to one year from date of shipment when the Product is subjected to normal, proper and intended usage. This warranty does not extend to anyone other than Buyer. Any model or sample furnished to Buyer is merely illustrative of the general type and quality of goods and does not represent that any Product will conform to such model or sample.
NO OTHER WARRANTIES, EXPRESS OR IMPLIED, ARE GRANTED, INCLUDING WITHOUT LIMITATION, IMPLIED WARRANTIES OF
MERCHANTABILITY, FITNESS FOR ANY PARTICULAR PURPOSE, OR NON INFRINGEMENT. BUYER’S EXCLUSIVE REMEDY FOR NON-
CONFORMING PRODUCTS DURING THE WARRANTY PERIOD IS LIMITED TO REPAIR, REPLACEMENT OF OR REFUND FOR THE NON-
CONFORMING PRODUCT(S) AT SELLER’S SOLE OPTION. THERE IS NO OBLIGATION TO REPAIR, REPLACE OR REFUND FOR PRODUCTS
AS THE RESULT OF (I) ACCIDENT, DISASTER OR EVENT OF FORCE MAJEURE, (II) MISUSE, FAULT OR NEGLIGENCE OF OR BY BUYER,
(III) USE OF THE PRODUCTS IN A MANNER FOR WHICH THEY WERE NOT DESIGNED, OR (IV) IMPROPER STORAGE AND HANDLING OF
THE PRODUCTS.
Unless otherwise expressly stated on the Product or in the documentation accompanying the Product, the Product is intended for research only and is not to be used for any other purpose, including without limitation, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses, or any type of consumption by or application to humans or animals.
Current product instructions are available at www.thermoscientific.com/pierce . For a faxed copy, call 800-874-3723 or contact your local distributor.
© 2012 Thermo Fisher Scientific Inc. All rights reserved. Unless otherwise indicated, all trademarks are property of Thermo Fisher Scientific Inc. and its subsidiaries. Printed in the USA.
Pierce Biotechnology
3747 N. Meridian Road
PO Box 117
Rockford, lL 61105 USA
3
(815) 968-0747
(815) 968-7316 fax www.thermoscientific.com/pierce

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