Update profile
    Change password
    Sign out

Pierce Abundant Protein Depletion Spin Columns


Deplete abundant plasma proteins quickly and economically.

Pierce Abundant Protein Depletion Spin Columns

Thermo Scientific Pierce Abundant Protein Depletion Spin Columns are optimized to decrease the abundant albumin and antibody components of human plasma samples in preparation for mass spectrometry or 1D and 2D gel electrophoresis.

The Pierce Top 2 Abundant Protein Depletion Spin Columns use highly specific immobilized anti-HSA and anti-IgG antibodies to remove human serum albumin (HSA) and all major subclasses of gamma globulin (IgG) from serum, plasma or spinal fluids. Similarly, the Pierce Top 12 Abundant Protein Depletion Spin Columns are designed to remove HSA, IgG and 10 other high-abundance proteins from human serum or plasma. The resins are provided in an economical and convenient spin column specifically designed one step processing and for single use.

Highlights:

  • Optimized – resin in spin columns is scaled and optimized for treating 10μL (600μg) of human plasma samples for mass spectrometry and/or 1D and 2D electrophoresis
  • Efficient – kits remove >90% of IgG and >95% of albumin, plus other abundant proteins (Top 12)
  • Fast – process samples in 40 to 60 minutes (depending on resin)
  • Economical – cost effective spin columns are priced for single use
  • Reduce variability – One time use prevents abundant protein carryover and experimental variability
  • Two resin options – choose the system appropriate for your need: 2 or 12 protein depletion columns
Video of Pierce Top 12 Abundant Protein Depletion Spin Columns in action!

Proteins removed by Pierce Abundant Protein Depletion Spin Columns. Binding and removal of proteins is achieved via specific antibodies, which are immobilized on the affinity support.
Top 2 Columns Top 12 Columns
  • Albumin
  • IgG
  • Albumin
  • IgG
  • α1-Acid Glycoprotein
  • α1-Antitrypsin
  • α2-Macroglobulin
  • Apolipoprotein A-I
  • Apolipoprotein A-II
  • Fibrinogen
  • Haptoglobin
  • IgA
  • IgM
  • Transferrin

Product Details:

Analysis of human fluids is often complicated by the presence of high concentrations of albumin and IgG that can make up more than 70% of total serum protein. Removal of these proteins is often essential for the study of low-abundant proteins. The Pierce Top 2 Protein Depletion Columns can deplete greater than 99% of HSA and 90% of IgG. The Pierce Top 12 Protein Depletion Columns can deplete greater than 95% of the twelve most abundant proteins in serum. Samples are loaded in a pre-filled convenient disposable spin format and can be processed in 40 to 60 minutes. The depletion of highly abundant proteins enables the detection of low abundant proteins in samples and subsequent identification by mass spectrometry or 1D and 2D gel electrophoresis.

Performance comparison of abundant protein depletion spin columns
Performance of Thermo Scientific Pierce Top 12 Abundant Protein Depletion Spin Columns compared to equivalent products from other suppliers. Human serum (10-20μL, Part No. 31876) was loaded onto each resin and processed according to the manufacturer’s protocol (Agilent: Human 14 Multiple Affinity Removal Spin Cartridge, Part No. 51886560; Sigma: SEPPRO* IgY14 spin column, Part No. SEP010-1Kit; BAC: CaptureSelect* HumanPlasma 14 Depletion Resin, Part No. 991410) protocol. Total protein in the depleted fractions was estimated using BCA protein Assay (Thermo Scientific, Part No. 23225). Total amount of albumin in the depleted fractions was determined using AssayMax* Human Albumin ELISA Kit (Assaypro, Part No. EA2201-1). FT = flow-through (i.e., depleted sample); E = eluate (i.e., proteins that were bound by the resin, plus stripped affinity antibodies of the column). Performance of all for products is comparable in this analysis. With the Top 12 proteins removed, low-abundance proteins are now visible in each depleted sample lane (FT).

 

Protein removal achieved using Pierce Top 12 Abundant Protein Depletion Spin Columns. Values were determined by targeted mass spectrometry. The albumin depletion percentage was cross-validated by ELISA and was in agreement with >99% removal.
Protein Fold Reduction Percent Depletion
albumin 3369 99.97
transferrin 266 99.62
alpha-1-antitrypsin 37 97.30
haptoglobin 127 99.21
alpha-1-acid glycoprotein 402 99.75
alpha-2-macroglobulin 116 99.14

 

Mass spectrometry peptide identification improved by abundant protein removal
Greater numbers of peptides identified following abundant protein removal. This proportional Venn diagram displays the relative number of unique peptides identified by mass spectrometry when human serum is depleted by the Thermo Scientific Pierce Top 2 or Top 12 Columns compared to non-depleted human serum. A simple, fast depletion using the Top 12 Columns doubled the number of unique peptides identified when compared to non-depleted human serum.

Cited References:

  1. Anderson, NL. and Anderson, NG. (2002). The human plasma proteome: history, character, and diagnostic prospects. Mol. Cell Proteomics. 1:845-67.
  2. Grundry, RL. et al. (2009). Assessment of albumin removal from an immunoaffinity spin column: Critical implications for proteomic examination of the albuminome and albumin-depleted samples. Proteomics. 9: 2021-8.

Related Resources:

Review of sample preparation for mass spectrometry

Related Products:

Albumin/IgG Removal Kit
Pierce Albumin Depletion Kit
Normal Human Serum
Protein Concentrators, 9K MWCO
Pierce Mass Spec Sample Prep Kit for Cultured Cells


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
85161 Top 2 Abundant Protein Depletion Spin Columns
Formulation: Microcentrifuge spin columns containing 0.6mL of affinity resin, supplied as a 62% slurry in PBS buffer (pH 7.4) with sodium azide
Sufficient For: Removing albumin and IgG from 6 human serum samples, 10μL each
6 columns Product Instructions for product #85161 Top 2 Abundant Protein Depletion Spin Columns MSDS for product #85161 Top 2 Abundant Protein Depletion Spin Columns Certificate of Analysis for product #85161 Top 2 Abundant Protein Depletion Spin Columns $154.00

Add to cart
85162 Top 2 Abundant Protein Depletion Spin Columns
Formulation: Microcentrifuge spin columns containing 0.6mL of affinity resin, supplied as a 62% slurry in PBS buffer (pH 7.4) with sodium azide
Sufficient For: Removing albumin and IgG from 24 human serum samples, 10μL each
24 columns Product Instructions for product #85162 Top 2 Abundant Protein Depletion Spin Columns MSDS for product #85162 Top 2 Abundant Protein Depletion Spin Columns Certificate of Analysis for product #85162 Top 2 Abundant Protein Depletion Spin Columns $436.00

Add to cart
85164 Top 12 Abundant Protein Depletion Spin Columns
Formulation: Microcentrifuge spin columns containing 0.81mL of affinity resin, supplied as a 62% slurry in PBS buffer (pH 7.4) with sodium azide
Sufficient For: Removing α1-acid glycoprotein fibrinogen, α1-anti-trypsin, haptoglobin, α2-macroglobulin IgA, albumin, IgG, apolipoprotein A-I, IgM, apolipoprotein A-II and transferrin from 6 human serum samples, 10μL each
6 columns Product Instructions for product #85164 Top 12 Abundant Protein Depletion Spin Columns MSDS for product #85164 Top 12 Abundant Protein Depletion Spin Columns Certificate of Analysis for product #85164 Top 12 Abundant Protein Depletion Spin Columns $200.00

Add to cart
85165 Top 12 Abundant Protein Depletion Spin Columns
Formulation: Microcentrifuge spin columns containing 0.81mL of affinity resin, supplied as a 62% slurry in PBS buffer (pH 7.4) with sodium azide
Sufficient For: Removing a1-acid glycoprotein fibrinogen, a1-anti-trypsin, haptoglobin, a2-macroglobulin IgA, albumin, IgG, apolipoprotein A-I, IgM, apolipoprotein A-II and transferrin from 24 human serum samples, 10┬ÁL each
24 columns Product Instructions for product #85165 Top 12 Abundant Protein Depletion Spin Columns MSDS for product #85165 Top 12 Abundant Protein Depletion Spin Columns Certificate of Analysis for product #85165 Top 12 Abundant Protein Depletion Spin Columns $610.00

Add to cart

Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

Share This Page
 
 

Follow Us
  Email Sign-up  Email Announcements

 



Antibodies  |   Molecular Biology   |   Cell Biology   |  Thermo Scientific  |   * Trademarks  |   Privacy Statement
© 2014 Thermo Fisher Scientific Inc.   |   3747 N Meridian Rd, Rockford, IL USA 61101   |   1-800-874-3723  or  815-968-0747