Protease inhibitor mixture method
Protease inhibitor mixture ( 100 ×, for mammalian sample extraction)
â— Storage conditions
Store at -20 ° C, valid for one year.
◠Product introduction <br> Protease inhibitor cocktail for mammalian cell and tissue extracts (100×) is a protease inhibitor for mammalian cell or tissue protein extraction. Mixtures (100 mMAEBSF, 80 μM Aprotinin, 5mMBestatin, 1.5mME64, 2mM Leupeptin and 1mMPepstatin A in DMSO) and provide individually packaged 0.5M EDTA.
Mammalian cells or tissue extracts contain many endogenous proteases, phosphatases, etc., which tend to cause degradation or de-modification of proteins in the extract, thereby affecting subsequent protein detection. Therefore, the addition of an appropriate protease, phosphatase or the like to the extract is an effective method for preventing protein degradation and de-modification.
This product is an optimized and tested mixture of protease inhibitors for mammalian cell or tissue protein extraction, containing a broad spectrum of serine, cysteine ​​and acid protease inhibitors, and aminopeptidase inhibitors.
This product is easy to use. Usually, the protease inhibitor mixture and 0.5M EDTA are added to the lysate at a ratio of 1:100. For the detection of metalloproteinase activity, EDTA should not be added.
The product can effectively inhibit various protease activities in mammalian cells or tissue extracts, such as serine protease, aminopeptidase, cysteine ​​protease, threonine and aspartic protease, metalloproteinase and the like. Among them, AEBSF is a water-soluble irreversible inhibitor of serine protease, and its inhibition constant is similar to the inhibition constant of PMSF and DFP, which can effectively inhibit trypsin, chymotrypsin, plasmin and kinin. Protease such as kallikrein and thrombin is used as a substitute for PMSF and DFP. AEBSF is less toxic, more water soluble and more stable in aqueous solution. Aprotinin is a competitive reversible inhibitor of serine protease; Bestatin is a reversible inhibitor of aminopeptidase; E64 is an irreversible inhibitor of cysteine ​​protease; Leupeptin is a reversible inhibitor of serine and cysteine ​​protease; Pepatatin A is aspartame Reversible inhibitor of acid protease; EDTA as a chelating agent is a reversible inhibitor of metalloproteinases.
A packaged product is usually sufficient for the formulation of 100 ml of lysate.
â— How to use:
1. The protease inhibitor mixture is a 100× stock solution, which is added to the lysate at a ratio of 1:100 (for example, 10 μl of protease inhibitor mixture is added to 1 ml of the lysate), and then mixed and used. base on needs,
0.5 M EDTA was also added to the lysate at a ratio of 1:100 (eg, for the detection of metalloproteinase activity, EDTA should not be added). The lysate containing the protease inhibitor mixture should be ready for use, and should not be formulated and stored for later use.
2. After the desired inhibitor has been added and mixed, the lysis and protein extraction of the mammalian tissue can be started.
Tip : Protease inhibitor mixtures should avoid repeated freezing and thawing. After the first use, they can be stored frozen according to the amount of each use.
Family: Arial'> 1:100 ratio is added to the lysate. 2. After the desired inhibitor has been added and mixed, the lysis and protein extraction of conventional cells or tissues can be started.
Tip : Protease inhibitor mixtures should avoid repeated freezing and thawing. After the first use, they can be stored frozen according to the amount of each use.
| Product number | specification | Storage | transport |
| PC2130-01 | 1 ml | -20 °C | Wet ice |
| PC2130-02 | 5 × 1 ml | -20 °C | Wet ice |
Store at -20 ° C, valid for one year.
◠Product introduction <br> Protease inhibitor cocktail for mammalian cell and tissue extracts (100×) is a protease inhibitor for mammalian cell or tissue protein extraction. Mixtures (100 mMAEBSF, 80 μM Aprotinin, 5mMBestatin, 1.5mME64, 2mM Leupeptin and 1mMPepstatin A in DMSO) and provide individually packaged 0.5M EDTA.
Mammalian cells or tissue extracts contain many endogenous proteases, phosphatases, etc., which tend to cause degradation or de-modification of proteins in the extract, thereby affecting subsequent protein detection. Therefore, the addition of an appropriate protease, phosphatase or the like to the extract is an effective method for preventing protein degradation and de-modification.
This product is an optimized and tested mixture of protease inhibitors for mammalian cell or tissue protein extraction, containing a broad spectrum of serine, cysteine ​​and acid protease inhibitors, and aminopeptidase inhibitors.
This product is easy to use. Usually, the protease inhibitor mixture and 0.5M EDTA are added to the lysate at a ratio of 1:100. For the detection of metalloproteinase activity, EDTA should not be added.
The product can effectively inhibit various protease activities in mammalian cells or tissue extracts, such as serine protease, aminopeptidase, cysteine ​​protease, threonine and aspartic protease, metalloproteinase and the like. Among them, AEBSF is a water-soluble irreversible inhibitor of serine protease, and its inhibition constant is similar to the inhibition constant of PMSF and DFP, which can effectively inhibit trypsin, chymotrypsin, plasmin and kinin. Protease such as kallikrein and thrombin is used as a substitute for PMSF and DFP. AEBSF is less toxic, more water soluble and more stable in aqueous solution. Aprotinin is a competitive reversible inhibitor of serine protease; Bestatin is a reversible inhibitor of aminopeptidase; E64 is an irreversible inhibitor of cysteine ​​protease; Leupeptin is a reversible inhibitor of serine and cysteine ​​protease; Pepatatin A is aspartame Reversible inhibitor of acid protease; EDTA as a chelating agent is a reversible inhibitor of metalloproteinases.
A packaged product is usually sufficient for the formulation of 100 ml of lysate.
â— How to use:
1. The protease inhibitor mixture is a 100× stock solution, which is added to the lysate at a ratio of 1:100 (for example, 10 μl of protease inhibitor mixture is added to 1 ml of the lysate), and then mixed and used. base on needs,
0.5 M EDTA was also added to the lysate at a ratio of 1:100 (eg, for the detection of metalloproteinase activity, EDTA should not be added). The lysate containing the protease inhibitor mixture should be ready for use, and should not be formulated and stored for later use.
2. After the desired inhibitor has been added and mixed, the lysis and protein extraction of the mammalian tissue can be started.
Tip : Protease inhibitor mixtures should avoid repeated freezing and thawing. After the first use, they can be stored frozen according to the amount of each use.
Family: Arial'> 1:100 ratio is added to the lysate. 2. After the desired inhibitor has been added and mixed, the lysis and protein extraction of conventional cells or tissues can be started.
Tip : Protease inhibitor mixtures should avoid repeated freezing and thawing. After the first use, they can be stored frozen according to the amount of each use.
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