Protein Stabilization Reagents. Elevated Protein Protection

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1 Protein Stabilization Reagents Elevated Protein Protection

2 Protein Stabilization Reagents Naturally occurring proteases and phosphatases can destroy proteins you spent days isolating. Save valuable time and resources purifying intact and functional protein with high yield. Choose the best product for your special application. Achieve broad-spectrum protection using convenient tablets from Roche. Ultra efficient protein protection To keep your proteins safe and intact, Roche has improved the protease inhibition capabilities of its proven cocktail tablets. Use the next-generation Tablets for even more effective protein protection in various lysates. Simultaneously preserve the original phosphorylation with PhosSTOP Phosphatase Inhibitor Cocktail Tablets. Proven protein protection Benefit from more than 18 years of Roche experience in protein isolation and protein protection against degradation with complete Tablets. Add the tablets directly to your lysate or prepare a stock solution with high concentration (up to 25x) and long stability (up to 3 months). Roche also offers a wide range of specific protease inhibitors for individual applications (see Ensure reliability Skip weighing out or coming into contact with hazardous protease inhibitor compounds. Use Roche Inhibitor Cocktail Tablets for reliable protein protection and improved laboratory safety. Protease Inhibitor Selection Guide Performance Convenience Tablets, EDTA-free Tablets complete Tablets, EDTA-free Serine Proteases Cysteine Proteases Aspartic Proteases Metalloproteases Stability stock solution Concentration stock solution* complete Tablets Table 1: or complete Tablets selection guide. (*Data refers to packaging for 50 ml; see ordering information).

3 Protease Inhibitor Cocktail Tablets Ideal for Cell Lysates Tablets were developed by optimizing their inhibitory effects on lysates from yeast, E. coli, and several mammalian cell lines (e.g., CHO-K1; see Figure 1). These tablets were shown to provide high level inhibition of serine, cysteine, aspartic, and metalloproteases (e.g., proteinase K; see Figure 2). 100 Inhibition of proteases in CHO-K Inhibition of proteinase K Inhibition ratio in % Inhibition ratio in % Tablets Supplier T Tablet P Supplier S Tablet S 0 Tablets Supplier T Tablet P Supplier T Liquid H Supplier S Tablet S Figure 1: Inhibition of proteases in CHO-K 1 lysate by complete ULTRA Tablets with EDTA in relation to other inhibitor tablets. Figure 2: Inhibition ratio of proteinase K by Tablets with EDTA in relation to other inhibitor cocktails. complete Lysis Kit Perform easier and faster cell lysis than traditional methods such as freeze-thaw cycles, sonicators, and glass beads. At the same time that cells are lysed, a multitude of proteases are inhibited by simply choosing either the complete Lysis Kit for bacteria or mammalian cells. For bacteria, use the complete Lysis-B (2x) Kit to efficiently and gently extract proteins in small volumes from bacteria (e.g., E. coli) and simultaneously inhibit proteases in the lysate. kd / complete Lysis-B Other reagent M W S P W S P Figure 3: Comparison of bacterial lysis reagents. W Whole fraction S Supernatant fraction P Pellet fraction GFP For life science research only. Not for use in diagnostic procedures. COMPLETE and PHOSSTOP are trademarks of Roche. All other brands or product names are trademarks of their respective holders.

4 Phosphatase Inhibitor Cocktail Tablets Performance of PhosSTOP Tablets Quickly and effectively inhibit protein dephosphorylation in your in vitro studies; simply drop a PhosSTOP Tablet into 10 ml buffer. Typical experiment with insect cells Lanes Inhibit a broad spectrum of phosphatase types, including acid, alkaline, serine/threonine, tyrosine, and dual-specificity phosphatases (see Table 2). Instantly inhibit phosphatases in a variety of sample materials, including mammalian, insect, or plant cells, as well as formalin-fixed, paraffin-embedded (FFPE) tissue sections. Freeze your phosphorylation status Phosphatases Units (U/10 ml) % Inhibition Calf alkaline phosphatase Potato acidic phosphatase 140 U 98.4% 2 U 93.7% Figure 4: Western blot of an insect cell lysate. Lanes 1 3: incubation without PhosSTOP Inhibitor for 0, 3, and 24 hours at +15 C to + 25 C. Lanes 4 6: incubation with PhosSTOP for 0, 3, and 24 hours at +15 C to + 25 C. Development of western blot: blocking with 1x solution of Western Blocking Reagent (Roche) in TBST, incubation with antiphosphoserine/threonine antibody (BD Bioscience) in blocking solution. Incubation with secondary antibody (Anti Mouse-Ig, Chemicon) in blocking solution. Detection using LumiLight PLUS Western Blotting Substrate (Roche). Human acidic phosphatase 640 U 99.5% Rabbit PP1 200 U 98.6% Human PP2A 500 U 94.4% Human PTP 500 U 96.7% Combination of PhosSTOP and complete or Tablets Inhibit both proteases and phosphatases at the same time. Simply use a complete UlTRA or complete Tablets and a PhosSTOP Tablet in the same buffer. Table 2: Effectively inhibit a broad range of phosphatases with a single PhosSTOP Tablet. After a 15-minute incubation (1 tablet per 10 ml), the inhibitory efficiency of the PhosSTOP Tablet was evaluated for alkaline (AP) and acid (SP) phosphatases, as well as for serine/threonine (PP1 and PP2A) and tyrosine protein phosphatases. For life science research only. Not for use in diagnostic procedures.

5 Ordering Information Product Catalog Number Pack Size Tablets, Mini, EDTA-free, EASYpack tablets in foil blisters (for 10 ml each) Tablets, EDTA-free, glass vials tablets in glass vials (for 50 ml each) 6 10 tablets in glass vials (for 50 ml each) Tablets, Mini, with EDTA, EASYpack tablets in foil blisters (for 10 ml each) Tablets, with EDTA, glass vials tablets in glass vials (for 50 ml each) 6 10 tablets in glass vials (for 50 ml each) complete Tablets, Mini, EDTA-free, EASYpack tablets in foil blisters (for 10 ml each) complete Mini tablets in glass vials (for 10 ml each) complete EDTA-free tablets in glass vials (for 50 ml each) 3 20 tablets in glass vials (for 50 ml each) complete Mini, EASYpack tablets in foil blisters (for 10 ml each) complete Mini, EDTA-free tablets in glass vials (for 10 ml each) complete tablets in glass vials (for 50 ml each) 3 20 tablets in glass vials (for 50 ml each) complete EDTA-free, EASYpack complete Lysis-B (2x), EDTA-free tablets in foil blisters (for 50 ml each) kit (100 ml (2x) lysis reagent/20 tablets) complete Lysis-M kit (200 ml (2x) lysis reagent/20 tablets) complete Lysis-M, EDTA-free kit (200 ml (2x) lysis reagent/20 tablets) PhosSTOP Tablets, EASYpack tablets (for 10 ml each) 20 tablets (for 10 ml each) Additional products: Roche also supplies single protease inhibitors for your selection for specific applications. See

6 Published by Roche Diagnostics GmbH Sandhofer Straße Mannheim Germany 2013 Roche Diagnostics. All rights reserved

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