Description :
Sunshinebio Protease and Phosphatase Inhibitor Mini Tablets, 2 tablets in vials, each tablet is sufficient for 10mL of extract
Storage: Upon receipt store at 4
Introduction The Sunshinebio Protease and Phosphatase Inhibitor Tablets protect proteins from degradation by endogenous proteases and phosphatases released during protein extraction and purification. Each tablet contains a mixture of several potent inhibitors (Table 1). The protease inhibitors target aminopeptidases and cysteine and serine proteases. The phosphatase inhibitors target serine, threonine and protein tyrosine phosphatases.
Table 1. Protease and phosphatase inhibitors included in the tablet. |
Inhibitor | Target Protease/Phosphatase |
|
Aprotinin | Serine proteases |
|
Bestatin | Aminopeptidase proteases |
|
E-64 | Cysteine proteases |
|
Leupeptin | Serine and cysteine proteases |
|
Sodium fluoride | Serine and threonine phosphatases |
|
Sodium orthovanadate | Tyrosine phosphatases |
|
Sodium pyrophosphate | Serine and threonine phosphatases |
|
β-glycerophosphate | Serine and threonine phosphatases |
|
Important Product Information
• The inhibitor formulation is typically effective when used at a 1X final concentration; however, samples containing high levels of proteases or phosphatases might require a more concentrated treatment (i.e., 2-3X).
• EDTA inhibits metalloproteases by chelating divalent cations necessary for their activity. By this same mechanism, EDTA can affect the activities of other proteins; empirically determine if EDTA is beneficial.
• The inhibitor formulation is compatible with most protein assays
• Pepstatin A is not included in the cocktail formulation; if you need to inhibit acid proteases, purchase Pepstatin A and add it to the solution.
• The inhibitor formulation interferes with immobilized metal-chelate affinity chromatography (IMAC) and 2D gel electrophoresis. Dialyze or desalt samples to effectively remove inhibitors from sample extracts before performing these procedures.
Procedure for using the Sunshinebio Protease and Phosphatase Inhibitor Tablet
1. Dispense from vial and place into 10mL of solution.
2. Allow the tablet to disperse in liquid for 30 seconds.
3. Vortex to dissolve; solution will be slightly hazy when tablet is dissolved.
Troubleshooting
Problem | Possible Cause | Solution |
Poor inhibition of protease or phosphatase activity | Sample contains high levels of proteases or phosphatases | Dissolve the tablet to 2X or 3X final concentration |
Tablet does not provide sufficient protection against a particular protease or phosphatase family | Inhibitor for a specific family was not a cocktail component | Add individual inhibitors to the samples for the specific protease or phosphatase |
A specific inhibitor was not at the needed concentration | Dissolve the tablet at a greater concentration |
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