Deplete abundant plasma proteins quickly and economically.
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
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 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
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:
Anderson, NL. and Anderson, NG. (2002). The human plasma proteome: history, character, and diagnostic prospects. Mol. Cell Proteomics. 1:845-67.
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.
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
$150.00
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
$425.00
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
$195.00
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 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
