ProteaPrep Albumin Depletion Sample Prep Kit

overview | features and specs | data sets | literature

Product Overview

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One of the major problems faced in the analysis of the human serum proteome is the broad dynamic range of its protein constituents. High abundance proteins, such as human serum albumin (HSA) and immunoglobulin gamma (IgG), which together comprise ~75% of the total serum protein content, inhibit the analysis of lower abundance proteins of interest.

Affinity-based depletion strategies are often employed to selectively remove high abundance proteins, such as HSA and IgG, to concentrate the lower abundance proteins and facilitate their analysis. Traditionally, antibody-based schemes are used to produce affinity-binding ligands for HSA and IgG, where monoclonal or polyclonal antibodies are produced in a source animal, purified, and then bound to a surface (e.g. bead or well) for preparation of serum samples. Variability in the production of these antibodies leads to a wide variety of specificities and selectivities, as the binding of the ligand to the HSA or IgG is dependent upon the availability of the epitope, the binding affinity of the antibody, and the homogeneity of the antibody population. Thus, the efficiencies for antibody-based depletion strategies for the selective removal of high abundance proteins in human serum can vary broadly, from 70 to 95%, and can dramatically affect the reproducibility of studies of the human serum proteome.

The ProteaPrep Albumin Depletion Sample Prep Kit (SP-200) features a revolutionary new approach to affinity-based serum albumin removal that utilizes a non-antibody-based method. The ProteaPrep Albumin Depletion capture ligand is a recombinant protein that exhibits excellent batch-to-batch reproducibility and is highly purified for robust, efficient removal of albumin from human serum samples. This proprietary ligand design technology produces binding constants with human serum albumin (Kd ~1 E-11M) that are significantly stronger than antibody-based methods, which have binding constants that range from Kd's of 1 E-6 to 1 E-8M. The net result is that the functionalized beads irreversibly bind and remove the albumin from serum in a highly efficient and rapid manner. Depletion efficiencies are >99% for the selective removal of HSA from serum samples in less than 20 minutes. The ProteaPrep Albumin Depletion Sample Prep Kit is also effective for the removal of human, rat, mouse, bovine, pig, goat, horse, rabbit, monkey and dog serum albumin.



Click images below to view data sets:

1) Single Image 2) Single Image 3) Single Image 4) Single Image
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1. SDS-PAGE gel showing depletion efficiency for HSA in human serum
2. Western blot showing depletion efficiency for standard HSA samples
3. Western blot showing depletion efficiency for HSA in human serum
4. SDS-PAGE gel showing mouse serum albumin standard depletion
5. SDS-PAGE gel showing depletion efficiency for standard HSA samples
6. SDS-PAGE gel showing rat serum albumin standard depletion
7. SDS-PAGE gel comparing ProteaPrep Albumin Depletion Sample Prep Kit to other common albumin depletion methods (data for courtesy of the Van Eyk Research Group at Johns Hopkins University, Baltimore, MD)
Features and Specifications
Features:
  • >99% efficiency for depletion of human serum albumin
  • Simple and easy to use - process samples in less than 20 minutes
  • New recombinant affinity ligand permits highly specific and reproducible albumin removal to facilitate analysis of lower abundance serum proteins
  • Albumin binding affinity 2 to 3 orders of magnitude greater than traditional antibody-based albumin depletion methods
  • Disposable, cost-effective SpinTube design eliminates the risk of carryover or sample contamination

Research Applications:
  • Human serum proteome analysis
  • Depletion of albumin from serum, plasma, and cell culture media
  • Preparation of serum samples for analysis by SDS-PAGE, 2D gel electrophoresis, and mass spectrometry

Specifications:
Proprietary recombinant affinity ligand for human serum albumin is covalently bound to 10µm spherical silica beads with 300Å pores.

This product is for R and D use only, not for drug, household, or other uses.
Data Sets

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Literature
***Please to download documentation.

ProteaPrep Albumin Depletion Sample Prep Kit Availability
This product is discontinued and no longer for sale.

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