Proteoprep® Kits

Depletes HSA and IgG from serum or plasma using small recombinant immunoaffinity ligands

Features & Benefits

• Novel antibody affinity media - Displays higher specificity and significantly higher binding capacity than conventional antibody-based resin kits.
• Reduces albumin and IgG contamination of spots - Improves 2-dimensional electrophoresis and later MALDI identification.
• More low abundance proteins - Increased loading (4-5 fold) volume of serum onto gel so that lower copy number proteins can be visualized and identified provide improved IEF resolution.
• Buffers are directly compatible with 2DE - Eliminating the need for protein precipitation procedures prior to electrophoresis.

back to top

For the sequential isolation of separate soluble cytoplasmic & membrane protein fractions

This kit features new and innovative detergents, and uses specially formulated reagents and an optimized protocol designed to generate two prepared subcellular fractions that are uniquely ready for two-dimensional (2D) electrophoresis. Fraction 1: Soluble/Cytoplasmic Proteins Fraction 2: Membrane Proteins The special reducing & alkylating reagents produce samples that exhibit improved focusing and decreased streaking in 2D gels. This kit provides reagents sufficient to process a minimum of ten samples, yielding two fractions each. This kit is appropriate for use with various model organism sample sources used in proteomics research. (Superior results produced from E. coli extractions.) Improved solubility allows for higher protein loading capacities, resulting in improved visualization of low abundance proteins in 2D gels.

Features & Benefits

• Innovative detergent preparations - Highly improved solubility allows for higher protein loads and greater visibility of low abundance proteins on 2D gels.
• Two pre-mixed solubilization solutions - Generates two distinct populations for easy 2D analysis.
• Pre-measured reducing and alkylating reagents - Easy-to-use reagents provide improved IEF resolution.
• Pre-weighed dry blends - Stable and easy to reconstitute.
• Conveniently Packaged - No waste. Use only the amount needed.

Components
  Soluble Cytoplasmic Extraction Reagent
  Soluble Protein Resuspension Reagent
  Cellular & Organelle Membrane Solubilizing Reagent
  Tributylphosphine Stock Solution
  Alkylating Reagent, Iodoacetamide

back to top

For total extraction of membrane proteins

This kit features new and innovative detergents, and uses specially formulated reagents and an optimized protocol to generate one fraction containing membrane proteins that is uniquely ready for two-dimensional (2D) electrophoresis. The special reducing and alkylating reagents produce samples that exhibit improved focusing and decreased streaking in 2D gels. This kit is appropriate for use with various model organism sample sources used in proteomics research. (Superior results produced from E. coli extractions.) Higher protein loading capacities and improved solubility, especially for difficult membrane bound proteins, provide excellent visualization of low abundance/low copy proteins.

Features & Benefits

• Innovative detergent preparations - Improved solubility allows for higher protein loads and greater visibility of low abundance proteins in 2D gels.
• Two pre-mixed solubilization solutions - Removes interfering non-membrane proteins prior to extraction, resulting in uncluttered 2D arrays.
• Pre-measured reducing & alkylating reagents - Easy-to-use reagents provide improved IEF resolution.

Components
  Soluble Cytoplasmic & Loosely-Bound Membrane Protein Extraction Reagent
  Cellular & Organelle Membrane Solubilizing Reagent
  Tributylphosphine Stock Solution
  Alkylating Reagent, Iodoacetamide

back to top

For testing or optimizing extraction conditions to produce total protein extracts

This kit provides four extraction reagents of increasing solubilizing power (reagents 1-4), each of which can generate total protein extracts from cellular samples. Along with conventional reagents, the kit also includes the newest generation of detergent reagents. This allows comparison of the protein extractions obtained with each of the four reagents and optimization to meet your individual needs. The reducing and alkylating reagents produce protein samples that exhibit improved focusing and decreased streaking in 2D gels. Enough of each component is provided to process a minimum of ten samples by each extraction reagent. For researchers who have optimized an extraction protocol using one chaotropic extraction reagent, each reagent is available as an individual product as well. (Superior results produced from E. coli extractions.)

Features & Benefits

• Four pre-mixed solubilization reagents - Enables rapid solublization.
• Pre-measured reducing and alkylating reagents - Easy-to-use reagents provide improved IEF resolution.
• Innovative detergent preparations - Highly improved solubility allows higher protein loads and greater visibility of low abundance proteins in 2D gels.

Components
  Chaotropic Membrane Extraction Reagent 1
  Chaotropic Membrane Extraction Reagent 2
  Chaotropic Membrane Extraction Reagent 3
  Cellular and Organelle Solubilizing Reagent 4
  Tributylphosphine Stock Solution
  Alkylating Reagent, Iodoacetamide

back to top

Albumin and IgG are two major protein components of serum representing ~80% of the total serum protein. Removal from Serum allows visualization of co-migrating proteins on a one-dimensional electrophoresis or two dimensional electrophoresis gel and higher sample load (4 to 5-fold) for improved visualization of lower copy number proteins (Figure 1). ProteoPrep Blue Albumin Depletion Kit specifically removes albumin and IgG from 25 samples of human serum (25 mL to 100 mL) in preparation for two-dimensional electrophoresis. The ProteoPrep Blue Albumin Depletion Medium is a mixture of two media: 1) a proprietary blue matrix and 2) Protein G agarose. Typical depletions are 85% for albumin and 70% for IgG from 75 mL of human serum. The medium exhibits low non-specific binding because it does not contain Cibacron® Blue, notorious for high non-specific binding of nonalbumin proteins. The kit reagents are urea-based buffers instead of salt-based buffers meaning the albumin-depleted serum samples can be applied in two-dimensional electrophoresis without protein precipitation. Focusing of proteins on immobilized pH gradient (IPG) strips for two-dimensional electrophoresis of proteins and mass spec analysis of in-gel digested spots is negatively impacted (e.g. poor spot resolution) by the presence of salts (Figure 2) or high concentrations of buffers. Many human serum albumin depletion products available today have salts in the equilibration and wash buffers typically requiring precipitation to remove the salts prior to two-dimensional electrophoresis which is time consuming and risks loss of important proteins. Experiments show that urea can replace salts allowing for albumin binding to a proprietary dye-based resin and inhibiting binding of non-albumin proteins.

Features & Benefits

• The ProteoPrep Blue media binds and removes 85% of the albumin and 70% of the IgG from 75 µl of serum allowing visualization of lower abundance proteins.
• Equilibration buffer, a Tris-buffered urea solution, is two-dimensional electrophoresis compatible and does not require sample precipitation.
• Compatible with sample sizes less than 25 µl without undesirable non-specific binding.

back to top

Customize & optimize your protein extraction using the most innovative detergents available.

Protein extraction is considered by many to be the most critical step of proteomic analysis; proteins to be studied must first be solublized. The ProteoPrep Detergent Sample pack contains 10 detergents, non-ionic and zwitterionic, for solubilization of membrane proteins. The variety of detergents enables testing and optimizing of extraction formulas, which vary with the protein of interest and cell type.

Detailed information on the Detergents

List of Components

back to top

The ProteoPrep Reduction and Alkylation Kit provides tributylphosphine (TBP) and iodoacetamide (IAA) for the reduction and alkylation of protein disulfide bonds. Procedures are provided for the reduction and alkylation of proteins in solution prior to loading the immobilized pH gradient (IPG) strip, during equilibration of the focused IPG strip prior to SDS-PAGE electrophoresis, or in-gel prior to tryptic digestion.

back to top

The ProteoPrep Protein Precipitation Kit contains separate trichloroacetic acid (TCA) and sodium deoxycholate (DOC) reagents for the precipitation of proteins from aqueous samples. TCA is a classic reagent for precipitating proteins; however, it is not effective in quantitatively precipitating proteins below the 30 µg level. The use of TCA in combination with DOC allows for precipitation of as little as 3 µg of protein. The kit provides reagents sufficient to precipitate fifty 0.1 mL protein samples.

Features & Benefits

• Fast - Precipitation in about 1 hour.
• Convenient - All 3 reagents are provided as ready-to-use solutions.
• Flexible - Resuspend pellet in the buffer of your choice to the protein concentration desired.
• Compatible - Choice of resuspension buffer offers compatibility with a number of downstream processes such as SDS-PAGE, protein assay, IEF, tryptic digest etc.
• Scalable - Procedures allow for precipitation to as little as 3 µg of protein.

Components
  Trichloroacetic Acid, 100% (w/v) Solution
  Deoxycholate, 0.2% (w/v) Solution
  Wash Solution, 25% (v/v) Acetone Solution

back to top
back to Extraction Kits & Reagents
back to Sample Preparation