Application

MS-SAFE Protease and Phosphatase Inhibitor has been used to measure ribosomal protein S6 kinase (S6K) activity.

Tested in mammalian cell lysates and liver tissue extracts. Designed for use in samples to be analyzed by mass spectrometry.

Components

Protease inhibitors: Bestatin hydrochloride Leupeptin Phosphoramidon disodium salt Pepstatin A Elastatinal Aprotinin Nafamostat mesylate AntipainPhosphatase inhibitors Okadaic acid Sodium fluoride Sodium orthovanadate Bromotetramisole oxalateFillers β-lactose DL-leucine

General description

Mass spectrometry (MS) compatible protease inhibitor cocktail and phosphatase inhibitor cocktail, with broad specificity for the inhibition of: Serine, cysteine and aspartic proteases, and metalloproteases Tyrosine, serine/threonine, acid and alkaline phosphatasesAll inhibitors and fillers were chosen so as not to interfere with LC-MS analyses. Any inhibitors capable of covalent, irreversible protein modification were avoided. This cocktail is EDTA-free. All inhibitors were selected to allow facile downstream sample processing, such as immobilized metal affinity chromatography (IMAC) for His-tagged protein purification and, uniquely, phosphopeptide enrichment.

Packaging

1 vial in glass bottle

Physical form

Lyophilized powder that is water-soluble

Quantity

One vial makes 20 mL of 1× inhibitor cocktail working solution, using either water or extraction/lysis buffer. Alternatively, a 10× concentrated solution may be prepared by adding 2 mL of water or extraction/lysis buffer. This 10× solution may then be diluted 10-fold into extraction/lysis buffer as needed for a 1× working solution.

Specificity

Protease inhibitors: serine, cysteine and aspartic proteases, and metalloproteasesPhosphatase inhibitors: tyrosine, serine/threonine, acid and alkaline phosphatases