ProteomeLab™ PA 800 Protein Characterization System--
Parts and Accessories

ProteomeLab PA 800 Protein Characterization System
The ProteomeLab PA 800 provides a comprehensive, automated and quantitative solution for the high-resolution characterization of proteins. The PA 800 application menu includes SDS-Gel Molecular Weight analysis, Isoelectric Focusing analysis, Peptide Mapping, Carbohydrate Profiling and the front-end separation and introduction of proteins to mass spectrometry. The high resolution analysis generated by the PA 800 provides more accurate information, and more precise quantitation yielding a better understanding of protein expression profiles.

The PA 800 System includes:

• High-resolution separation module
• High-sensitivity UV, PDA, and laser-induced fluorescence detection
• Sample storage temperature control
• High-speed system controller with integrated application-based data analysis
• Validated turnkey methods and system application guides
• Starter kit of necessary hardware supplies to operate the system
• Installation Qualification and Operation Qualification of the system
• System application installation and training

This part number must be ordered with 285600 for installation.

ProteomeLab PA 800s SDS-MW Analysis System
A subset of the ProteomeLab PA 800 Protein Characterization system, the ProteomeLab PA 800s provides an automated and quantitative solution for the analysis of protein-SDS complexes by their molecular weight.

The PA 800s system includes:

• High-resolution separation module
• High-sensitivity PDA detection
• Sample storage temperature control
• High-speed system controller with integrated application-based data analysis
• Validated turn-key SDS-MW method and system application guide
• Starter kit of necessary hardware supplies to operate the system
• Installation Qualification and Operation Qualification of the system
• System application installation and training

This part number must be ordered with 713360 for installation.

ProteomeLab PA 800 System Start-Up Chemistry
Supporting chemistry for the PA 800 application menu.

The Start-Up Chemistry includes:

• ProteomeLab PA 800 System Performance Test
• ProteomeLab SDS-Gel MW Analysis
• ProteomeLab Carbohydrate Labeling and Analysis
• ProteomeLab Isoelectric Focusing

ProteomeLab PA 800—Performance Test
The ProteomeLab PA 800 Performance Test contains calibration standards, test standards, and buffer systems for system performance evaluation.

This includes:

• ProteomeLab PA 800 System Suitability Test Standard and Buffers
• LIF Calibration Standard
• Integrated PA 800 - MS/MS Test Standard and Buffer
• ProteomeLab PA 800 Performance Test Guide

ProteomeLab SDS-Gel MW Analysis Chemistries
The ProteomeLab SDS-Gel MW Analysis is designed for the separation and sizing of protein–SDS complexes using a replaceable gel matrix. The gel is formulated to provide an effective protein sieving range of approximately 10 kDa to 225 kDa. Within this size range, the logarithm of protein molecular mass is linear with its reciprocal electrophoretic mobility, allowing the molecular weight of an unknown protein to be estimated from a standard curve of known protein sizes. This chemistry can also be used to effectively quantify the amount of protein and to determine the purity of a protein product.

This includes:

• Separation Capillary, 57 cm x 50µm ID bare fused silica, 2 pieces
• SDS Gel Separation Buffer, 140 mL
• SDS Sample Buffer, 100 mM Tris-HCl, pH 9.0/1% SDS, 50 mL
• SDS Protein Sizing Standard (10 to 225 kDa) 16 mg/mL, 100 µL
• Internal Standard, 10 kDa protein, 5 mg/mL, 0.4 mL
• Acidic Wash Solution, 0.1 N HCl, 100 mL
• Basic Wash Solution, 0.1 N NaOH, 100 mL
• SDS-Gel MW Analysis Guide, 1 piece

ProteomeLab IgG Purity/Heterogeneity Assay
Includes IgG Control Standard 1-pack and all chemistries.
The ProteomeLab IgG Purity/Heterogeneity Assay has been developed for researchers employed in industrial biotechnology, who are developing and manufacturing IgG reagents for research, diagnostic and therapeutic use. This assay has been specified to assess the purity and heterogeneity of IgG reagents in both a reduced and non-reduced state. The methodology involves heat denaturing a specified concetration of IgG (both reduced and non-reduced) in the presence of SDS, and separating these proteins by size using high resolution capillary gel electrophoresis technology. This assay will detect impurities as low as 0.1% and includes an IgG control with a designated quantity of non-glycosylated heavy chain to test both the resolution and quantitation suitability of the assay prior to running unknowns.

Assay chemistry includes:

• SDS gel buffer (140 mLs) - The gel buffer comprises a proprietary polymer buffer formulation at pH 8.0, with 0.2% SDS.
• SDS sample buffer (50 mLs) - The sample buffer is composed of 100 mM Tris-HCL pH 9.0 with 1% SDS.
• IgG Control Standard (1 mL) - A well characterized molecule with a controlled quantity of non-glycosylated heavy chain, providing both a resolution and quantification benchmark. In SDS sample buffer at a concentration of 1 mg/ml.
• 10 kDa Internal Standard - used as an internal standard in this assay. Packaged as a stock solution at a protein concentration of 5 mg/ml in 0.5% SDS, 0.2% sodium azide.
• Acidic wash solution (100 mLs) - a high purity reagent comprised of 0.1 N HCL, used to regenerate the capillary surface.
• Basic wash solution (100 mLs) - a high purity reagent comprised of 0.1 N NaOH used to regenerate the capillary surface.
• Standard Operating Protocol - detailed instructions to perform assay
• Separation Capillary (Qty 2) - 50 um id x 57 cm bare-fused silica capillary

Carbohydrate Labeling and Analysis Chemistries
The ProteomeLab PA 800 Carbohydrate Labeling & Analysis Chemistry contains the reagents, buffers and separation capillaries required to label, separate and quantify oligosaccharides and monosaccharides released from glycoproteins. After release (enzymatic or chemical), sugars are labeled with a fluorophore, APTS, at the reducing termini by reductive amination. The stoichiometry of labeling is such that only one APTS molecule is attached to each molecule of oligosaccharide. These highly charged and fluorescent oligosaccharides are easily resolved in an electric field and detected by laser-induced fluorescence.

This includes:

• Carbohydrate Separation Buffer, 56 mL
• N-CHO Coated Capillary, 2 pieces
• Labeling Reagents, 1 piece
  • Labeling Dye (APTS), 4 x 5 mg
  • Labeling Dye Solvent, 1 mL
  • Glucose Ladder Standard, 50 mg
  • Quantitation/Mobility Marker (Maltose), 0.18 mg
• APTS-M (monosaccharide grade), 20 mg
• Carbohydrate Labeling and Analysis Guide, 1 piece

Isoelectric Focusing Chemistries
Isoelectric focusing (IEF) has been widely used for the separation of proteins based on differences in their isoelectric points. The IEF method is accomplished by electrophoresis of proteins or peptides through a stable pH gradient until they reach the pH equal to their isoelectric point (pI), at which time the net charge and mobility of the protein are zero. The ProteomeLab PA 800 System automates this process with the use of online capillary isoelectric focusing. The applicable pI range of this capillary isoelectric focusing chemistry is pH 3.0-10.0. The migration time of the sample of interest may be correlated with the calibration plot of pI markers. In addition to providing separation with high resolution, this chemistry may be used for wide-range (pI 3-10) screening and pI identification of unknown proteins and peptides.

This includes:

• Neutral Separation Capillary, 50 µm ID x 45 cm, 1 piece
• cIEF 3-10 Amphohyte, 1 mL
• cIEF Gel (Polymer solution), 100 mL
• 1M Phosphoric Acid, 10 mL
• 1M Sodium Hydroxide, 3.1 mL
• Ribonuclease A (pI 9.45), 3.84 mL
• Carbonic Anhydrase II (pI 5.9), 0.32 mg
• Lactoglobulin A (pI 5.1), 0.64 mg
• CCK Flanking Peptide (pI 2.75), 0.16 mg
• Isoelectric Focusing Guide, 1 piece

Protein Methods Development Chemistries
The Protein Methods Development Chemistries contain coated capillaries, buffers, standards, and markers to allow you to optimize a separation method for the analysis of a broad spectrum of proteins by their mass/charge characteristics. The use of a "Neutral" separation capillary minimizes the adsorption of proteins to the capillary surface and protects against hydrophobic interactions with the surface, improving the overall efficiency and resolution of the proteins being separated.

This includes:

• Neutral Capillary, 50 µm, 1 piece
• Orange G Reference Marker, 0.1% aqueous solution, 1 mL
• Histamine Reference Marker, 0.1% aqueous solution, 1 mL
• Citrate Buffer, pH 3, 50 mM, 100 mL
• Citrate/MES Buffer, pH 6, 50 mM, 100 mL
• Tricine Buffer, pH 8, 50 mM, 100 mL
• Protein Test Mix, 1 piece
  • Lyzozyme, 1 mg
  • Ribonuclease A, 1 mg
  • Cytochrome C, 3 mg
• Methods Development Guide, 1 piece

Upgrade to ProteomeLab PA 800/PA 800s from MDQ with Software Version 4.0 or Greater
Eligible Systems: P/ACE™ MDQ with Sample Temperature control and PDA detection—SN 3060526 (July 1999) cutoff applies.

This upgrade includes:

• PA 800 Hardware Upgrade (firmware, vial ejection system, electronic updates, exterior)
• 32 Karat™ 7.0 Workstation configured for PA 800
• Validated turnkey SDS-MV method and system application guide
• PA 800s Shipkit
• Installation Qualification and Operator Qualification of the system
• System application installation and training

This part number must be ordered with 285600 or 713360 for installation.

Upgrade to ProteomeLab PA 800/PA 800s from MDQ with Software Version < 4.0
Eligible Systems: P/ACE™ MDQ with Sample Temperature control and PDA detection—SN 3060526 (July 1999) cutoff applies.

This upgrade includes:

• PA 800 Hardware Upgrade (firmware, vial ejection system, electronic updates, exterior)
• 32 Karat™ 7.0 Workstation configured for PA 800
• Validated turnkey SDS-MV method and system application guide
• PA 800s Shipkit
• Installation Qualification and Operator Qualification of the system
• System application installation and training
• 32 Karat version 7.0 license

This part number must be ordered with 285600 or 713360 for installation.