Roche Applied Science protein research techniques build on years of experience in protein labeling, detection, purification and stabilization. cOmplete products protect your proteins during cell lysis, by offering reliable and best-in-class protease inhibition and with full compatibility with EDTA and DTT when performing His-Tag Purification with IMAC.

Protease Inhibitors
 

> cOmplete ULTRA Protease Inhibitor Tablets cOmplete ULTRA Tablets are the most powerful protease inhibitor cocktail available. In addition to our well known and reliable cOmplete Tablets cOmplete ULTRA Tablets inhibit aspartic proteases for maximum protection. Please visit the selection table

> cOmplete Protease Inhibitor Tablets The cOmplete Tablets were the first inhibitor cocktail tablets on the market and invented by Roche Applied Science. They are available in all varieties (with and without EDTA; blister pack and glass vials) and a safe and well-known solution against protein degradation by proteases. For more than 10 years thousands of researchers have protected their proteins with cOmplete Tablets.

His-Tag Purification

> cOmplete His-Tag Purification Resin The cOmplete His-Tag Purification Resin is a high-capacity IMAC (Immobilized Metal Affinity Chromatography) matrix for convenient single-step purification of His-tagged proteins from lysates. Its nickel-chelate chemistry is truly compatible with EDTA and DTT, and a wide range of buffer substances and salt conditions. Nickel leakage is minimized, allowing for multiple reuse without recharging nickel ions without any loss of capacity or purity.

Cell Lysis 

> cOmplete Lysis Kits cOmplete Lysis Kits make cell lysis easier and faster than traditional methods such as freeze-thaw cycles, sonicators and glass beads. At the same time that cells are lysed, a multitude of proteases can be inhibited. Simply add the cOmplete Lysis Kits optimized for bacteria and mammalian cells.

Phosphatase Inhibition

> PhosSTOP Tablets PhosSTOP Phosphatase Inhibitor Cocktail Tablets are a proprietary, non-toxic blend of phosphatase inhibitors, which dissolve quickly in aqueous solutions (buffers) and work also in buffers containing formalin for formalin-fixation. PhosSTOP is formulated as a ready-to-use tablet since some of the phosphatase inhibitors are only necessary in nanomol amounts.

Related Products
 

Individual Protease Inhibitors

Cells contain different types of proteases. Therefore, usually more than one protease inhibitor is needed for complete preservation of the cell's protein(s) which shall be isolated for subsequent experiments, such as 2-D electrophoresis, mass spectrometry, liquid chromatography, specific protein-activity assays, or other experiments performed in proteome research.
 

• See full list of protease inhibitors here

Protein Processing
Highly purified and specific proteases are essential tools for proteome research. In particular proteins that will be analyzed by mass spectrometry usually must be cleaved after purification (e.g., by 2-D gel electrophoresis). A reproducible cleavage pattern is a prerequisite for clear identification. Trypsin and, to a smaller extent, other endoproteinases (endoproteinases Arg-C, Lys-C, Glu-C) are used to perform these cleavages.
Roche Applied Science offers a recombinant trypsin, proteomics grade, to specifically address these requirements. In addition, we offer different sequencing-grade endoproteinases suitable for the high-performance cleavages listed below
 

• Sequencing-Grade Proteases for Proteomics Research
• Other Proteases for Proteomics Research
   

Protein Characterization
Following identification of a specific gene product, the protein(s) must be further characterized. For instance, glycan analysis can show whether a protein went through the secretory pathway. For further functional characterization with regard to, for example, subcellular localization or expression patterns in different cell types, labeling and detection tools are required. Analysis of the protein of interest is done either by the generation of a corresponding antibody or by the introduction of an epitope tag, a short amino acid sequence. Furthermore, labeling of purified proteins or antibodies with biotin, digoxigenin, fluorescein, or enzymes, such as peroxidase (POD) or alkaline phosphatase (AP), enables their detection. Finally, methods such as Western blotting or ELISA are used for the qualitative or quantitative detection of certain proteins.
 

• Glycan Analysis
  Tools for the detection and characterization of carbohydrate modifications.
   
• Labeling of Proteins and Detection of Proteins
  Kits and reagents for the labeling of proteins with ligands, such as alkaline phoshatase, biotin, digoxigenin, fluorescein, or peroxidase, and for the detection of proteins that have been modified with such ligands.
   
• Epitope Tagging
  Antibodies for the detection of proteins carrying commonly used epitope sequences like the influenza hemagglutinin protein (HA), human c-myc protein, or six histidine residues (HIS₆).
   
• Western Blotting
  Everything you need for the detection of proteins by chemiluminescence or chromogenic methods following Western blotting.
   
• ELISA
  Kits and reagents for the detection of proteins using the ELISA technique.

> Protease Inhibitors Selection Guide

Protein Purification Selection Guides

 > Protein Purification via Tag
 > Protein Purification via Antibody



 

Protease Inhibitors Selection Guide

Use the following information to select the appropriate protease inhibitor for your application or see the full product list:
 

Protein Purification via Tag

Purification via Antibodies using Protein A, Protein G and Protein L

Use this table to select the appropirate product to purify antibodies from solutions

++= strong binding; += moderate binding; -=no binding

 

Ultra Efficient Protein Protection

In just minutes, proteases can destroy the proteins you have spent days isolating. Upgrade to the most powerful protease inhibitor cocktail available. cOmplete ULTRA Protease Inhibitor Cocktail Tablets secure your proteins against an unprecedented range of proteases.

cOmplete ULTRA Tablets contain a proprietary blend of protease inhibitors. Simply add a tablet to your homogenization buffer and instantly protect your proteins against a broad range of proteases. cOmplete ULTRA Tablets contain both irreversible and reversible protease inhibitors.

• Increase intact protein yield by efficiently inhibiting an even wider range of proteases using this improved protease inhibitor cocktail; acidic proteases and additional serine proteases are also inhibited.
• Ensure laboratory safety by avoiding contact with hazardous compounds, and deliver a consistent dose without the need to weigh out small amounts of inhibitors.
• Specifically formulated for cell lysates. cOmplete ULTRA Tablets have been developed using careful evaluation of inhibitory effects on lysates from yeast, E. coli, and insect cells, as well as from several mammalian cell lines (e.g., HEK-293, CHO, COS). These tablets provide excellent inhibition of serine, cysteine, and acidic proteases in all of those lysates.

Competitor comparison

Inhibition ratio of cOmplete ULTRA Tablets versus other protease inhibitor cocktail tablets with and without EDTA:
 

Figure 1: Inhibition ratio of proteases in CHO K1 lysate with cOmplete ULTRA Tablets with EDTA in relation to other tablet suppliers.	Figure 2: Inhibition ratio of proteases in CHO K1 lysate with cOmplete ULTRA Tablets without EDTA in relation to other tablet suppliers.

Request your free sample and discover how cOmplete ULTRA tablets can improve your yields of full-length protein.

Combine cOmplete ULTRA tablets with PhosSTOP Phosphatase Inhibitor Cocktail Tablets for simultaneous protection against proteases and phosphatases.

IMAC purification now truly compatible with EDTA and DTT

cOmplete His-Tag Purification Resin  is an innovative high-capacity IMAC matrix (Immobilized Metal Affinity Chromatography) for convenient single-step purifications of His-tagged proteins from total lysates. Roche's propriety nickel-chelate chemistry ensures extraordinary compatibility with commonly used reducing agents such as DTT, chelating metalloprotease inhibitors such as EDTA, and a wide range of buffer substances and salt conditions. The wide choice of compatible ingredients allows optimization of buffers for maximum protein stability and solubility.

cOmplete His-Tag Purification Resin is the only resin for purifying large amounts of protein without compromises.
 

• Use the buffer conditions best suited to your protein
  Keep your protein comfortable and let it, not your purification resin, determine whether you use DTT, EDTA, or other buffer substances.
• Repeatedly obtain highly pure protein
  Single step purification without resin recharging.
• Protect your protein from toxic nickel
  Reduce protein oxidation and aggregation caused by resins that leach nickel.
• Work in a safe and eco-friendly environment
  Avoid handling of toxic nickel and completely eliminate disposal costs
   

Figure 1: Tight Binding of nickel visualized. Before (left) and after (right) photos of cOmplete His-Tag Purification Resin after 5 times reuse without recharging.

 

 

Figure 2: Protein-binding performance with His6 CFP. cOmplete His-Tag Purification Resin (blue columns) with 10 mM DTT and EDTA is reused without nickel recharging alongside Resin G (grey columns) with 1 mM EDTA and 5 mM DTT ( as specified in manufactor's package insert). Another competing product, Resin Q (not shown), did not bind any protein at all.	Figure 3: Loss of resin Ni ions under stringent conditions. One milliliter each of cOmplete His-Tag Purification Resin and 2 commercially available resins were incubated in 9 ml of a buffer containing 50 mM NaH2PO4, 300 mM NaCl, pH 8.0, 10 mM EDTA, 10 mM DTT, and 500 mM imidazole. The cOmplete Resin lost less than 1 percent of nickel ions.

>  Request your free sample and discover how efficiently this new resin purifies your His-tagged proteins.