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Cat. No.: PPIC-10 (for 10ml each)

Description

Protease and phosphatase inhibitor cocktail for general use (50X) is a combination of protease and phosphatase inhibitor mixtures used for protein extraction from cells or tissues. It contains one bottle of protease inhibitor cocktail (general use, 50X) (100mM AEBSF, 15μM Aprotinin, 6.5mM Bestatin, 0.7mM E64, 0.5mM Leupeptin in DMSO) and one bottle of phosphatase inhibitor cocktail (50X) (250mM sodium fluoride, 50mM sodium pyrophosphate, 50mM β-glycerophosphate, and 50mM sodium orthovanadate in H2O), along with individually packaged 0.05M EDTA.

Cell or tissue extracts often contain endogenous proteases, phosphatases, and other enzymes that can lead to protein degradation or modifications, which can affect subsequent protein analysis. Therefore, adding appropriate protease and phosphatase inhibitors to the sample during extraction is an effective method to prevent protein degradation and modifications.

This product is an optimized and tested mixture of protease and phosphatase inhibitors for general cell or tissue protein extraction. It includes a broad-spectrum of serine, cysteine, and acid protease inhibitors/aminopeptidase inhibitors, as well as serine/threonine, tyrosine, acid, and alkaline phosphatase inhibitors.

The protease and phosphatase inhibitor cocktail contains 100mM AEBSF, 15μM Aprotinin, 6.5mM Bestatin, 0.7mM E64, 0.5mM Leupeptin, 250mM sodium fluoride, 50mM sodium pyrophosphate, 50mM β-glycerophosphate, and 50mM sodium orthovanadate, prepared in high-purity DMSO and ultrapure water. EDTA is separately packaged for selective use. If detecting metalloproteinase activity, EDTA should not be added.

This product is convenient to use. Typically, the protease inhibitor cocktail (general use, 50X), phosphatase inhibitor cocktail (50X), and 0.05M EDTA are added to the lysis buffer at a ratio of 1:50 for routine cell or tissue protein extraction, effectively inhibiting protein degradation.

Different types of protein extracts contain different types of proteases. For example, mammalian cell extracts typically contain high levels of serine proteases, bacterial cell extracts contain a higher amount of serine proteases and metalloproteinases, mammalian tissue extracts mainly contain serine, cysteine, and metalloproteinases, plant extracts often contain a large amount of serine and cysteine proteases, and some may contain aspartic proteases.

This product can effectively inhibit various proteases (such as serine proteases, cysteine proteases, aspartic proteases, metalloproteases, etc.) and phosphatases (such as serine/threonine phosphatases, tyrosine phosphatases, acid phosphatases, alkaline phosphatases, etc.) in conventional cell or tissue extracts.

AEBSF, a water-soluble irreversible inhibitor of serine proteases, has a similar inhibitory constant to PMSF and DFP. It can effectively inhibit proteases such as trypsin, chymotrypsin, plasmin, kallikrein, and thrombin. As a substitute for PMSF and DFP, AEBSF has lower toxicity, better water solubility, and solution stability.

Aprotinin is a competitive reversible inhibitor of serine proteases. Bestatin is a reversible inhibitor of aminopeptidases. E64 is an irreversible inhibitor of cysteine proteases. Leupeptin is a reversible inhibitor of serine and cysteine proteases. Pepstatin A is a reversible inhibitor of aspartic proteases. EDTA, as a chelating agent, is a reversible inhibitor of metalloproteases. Sodium fluoride is a reversible inhibitor of acid phosphatases. Sodium pyrophosphate is an irreversible inhibitor of serine/threonine phosphatases. β-glycerophosphate is a reversible inhibitor of serine/threonine phosphatases. Sodium orthovanadate is a reversible inhibitor of alkaline phosphatases and tyrosine phosphatases.

One package of this product is typically sufficient for the preparation of 500 ml of lysis buffer.

Precautions

• This product can be properly aliquoted and stored. It should be added to the lysis buffer or appropriate solution shortly before use.
• The product provides separately packaged EDTA, which can be selectively used. For example, if it is intended for the detection of metalloproteinase activity in the extraction solution, EDTA should not be added.
• The protease and phosphatase inhibitor cocktail contained in this product can be harmful to the human body. Please handle with care and take necessary precautions to avoid direct contact or inhalation.
• This product is intended for scientific research purposes by professionals only. It is not intended for clinical diagnosis or treatment, food or drug applications, and should not be stored in a residential setting.
• For your safety and health, please wear appropriate laboratory attire and disposable gloves when handling this product.

Storage

Store at -20°C, valid for one year.

Only for research and not intended for treatment of humans or animals

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