Complete protection from proteases and phosphatases in one reliable cocktail. Perform protein extractions and purifications with confidence.

Thermo Scientific Halt Protease and Phosphatase Inhibitor Cocktail provides the convenience of a single solution with full sample protection. This broad-spectrum inhibitor cocktail is specifically optimized to protect proteins from degradation during extraction and purification. The cocktail contains inhibitors against the major classes of proteases and phosphatases, targeting aminopeptidases, cysteine and serine proteases and serine/threonine and protein tyrosine phosphatases. To promote the inhibition of metalloproteases, 0.5 M EDTA is provided in a separate tube.

Highlights:

• Convenient all-in-one cocktail contains both protease and phosphatase inhibitors
• Four convenient formats are available: single-use, 1 ml, 5 x 1 ml and 10 ml
• Immediate inhibition - no need to thaw, open multiple tubes or wait for tablets to dissolve
• Compatible with Thermo Scientific Pierce Cell Lysis Reagents
• Compatible with mass spectrometry (MS)

This combined cocktail is the first of its kind. The aqueous-based format allows for convenient cold-room storage without freezing, which is associated with DMSO-based cocktails, and is much easier to use than tablets that require tedious splitting with a razor blade to treat samples < 10 ml (the minimum sample volume for tablets is 10 ml). More importantly, the cocktail formulation is more effective than tablet formats for inhibiting proteases in lysates and preserving protein phosphorylation during cell harvest. The formulation is also MS-compatible because it does not contain AEBSF, which can cause peaks to shift.

Thermo Scientific Halt Protease and Phosphatase Inhibitor Cocktails are more effective than tablet-format protease inhibitor cocktails. Using a general protease assay with a rat pancreas extract (1 mg/ml), the Halt Protease and Phosphatase Inhibitor Cocktails (± EDTA) were compared with the tabletbased protease inhibitor cocktails (± EDTA) of Competitor Tablet. The same experimental conditions were used for all samples tested. A 1X final concentration of each inhibitor cocktail was added to the extract. The Halt Protease and Phosphatase Inhibitor Cocktails resulted in  ≥ 97% inhibition compared to  ≥ 62% inhibition for the tablets.

Thermo Scientific Halt Protease and Phosphatase Inhibitor Cocktail preserves phosphorylation of phosphorylated PTEN and CDC2 in HeLa cell lysate more effectively than tablet-format phosphatase inhibitor cocktail. HeLa cells were serum-starved for 24 hours. Serum-containing media was added to the cells for 15 minutes and cells were lysed with Thermo Scientific M-PER Mammalian Protein Extraction Reagent (Product # 78501) in the presence or absence of 1X inhibitor cocktail. Tablets were dissolved in 1 ml of buffer and then diluted 1:100 into the lysis reagent. The Halt Protease and Phosphatase Inhibitor Cocktail was added directly to the lysis reagent. Lysates were analyzed by Western blot for preservation of protein phosphorylation using the antibodies indicated. Panel A. Phosphorylated PTEN, Panel B. Phosphorylated CDC2. The antibodies recognize phosphorylation on serine and tyrosine, respectively.

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