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Cat. No.: PPICMT-2 (for 2ml each)

Cat. No.: PPICMT-10 (for 10ml each)

Description

Protease and Phosphatase Inhibitor Cocktail (for Mammalian Cell and Tissue Extracts, 50X) is a protease and phosphatase inhibitor mixture for mammalian cell or tissue protein extraction. This product contains one tube of protease inhibitor (for mammalian sample extraction, 50X) (52.5mM AEBSF, 40μM Aprotinin, 2.5mM Bestatin, 0.75mM E64, 1mM Leupeptin, and 0.75mM Pepstatin A in DMSO) and one tube of phosphatase inhibitor (50X) (250mM sodium fluoride, 50mM sodium pyrophosphate, 50mM β-glycerophosphate, and 50mM sodium orthovanadate in H2O), and includes separately packaged 0.25M EDTA.

Mammalian cell or tissue extracts contain many endogenous proteases and phosphatases, which can easily lead to protein degradation or de-modification in the extracts, affecting subsequent protein detection. Therefore, adding appropriate protease and phosphatase inhibitors to extracts is an effective method to prevent protein degradation and de-modification.

This product is an optimized and tested protease and phosphatase inhibitor mixture for mammalian cell or tissue protein extraction, containing broad-spectrum serine, cysteine, and acidic protease inhibitors/aminopeptidase inhibitors, as well as serine/threonine, tyrosine, acidic, and alkaline phosphatase inhibitors.

This protease and phosphatase inhibitor mixture contains 52.5mM AEBSF, 40μM Aprotinin, 2.5mM Bestatin, 0.75mM E64, 1mM Leupeptin, 0.75mM Pepstatin A, 250mM sodium fluoride, 50mM sodium pyrophosphate, 50mM β-glycerophosphate, and 50mM sodium orthovanadate, formulated in high-purity DMSO and ultrapure water, respectively. EDTA is packaged separately for selective use. If detecting metalloprotease activity, EDTA should not be added.

This product is convenient to use. Typically, the protease inhibitor mixture (for mammalian sample extraction, 50X), phosphatase inhibitor (50X), and 0.25M EDTA are added to the lysis buffer at a ratio of 1:50, respectively, for the extraction of proteins from mammalian cells or tissues, effectively inhibiting protein degradation.

Different types of protein extracts contain different types of proteases. For example, mammalian cell extracts usually contain high levels of serine proteases, bacterial cell extracts often contain more serine and metalloproteases, mammalian tissue extracts mainly contain serine, cysteine, and metalloproteases, and plant extracts often contain large amounts of serine and cysteine proteases, with some containing aspartic proteases. Therefore, we provide different types of protease and phosphatase inhibitor mixtures for different sample types.

This product can effectively inhibit various protease activities (such as serine proteases, aminopeptidases, cysteine proteases, threonine and aspartic proteases, metalloproteases, etc.) and phosphatase activities (such as serine/threonine, tyrosine, acidic, and alkaline phosphatases) in mammalian cell or tissue extracts. AEBSF is a water-soluble, irreversible serine protease inhibitor with an inhibition constant similar to PMSF and DFP, effectively inhibiting proteases such as trypsin, chymotrypsin, plasmin, kallikrein, and thrombin. As a substitute for PMSF and DFP, AEBSF has lower toxicity, better water solubility, and stability in aqueous solutions. 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 acidic 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.

Storage

Store at -20°C for one year.

Precautions

• This product can be aliquoted and stored, and should be added to the lysis buffer or other appropriate solutions just before use.
• This product includes separately packaged EDTA for selective use. For example, if the extract will be used to detect metalloprotease activity, EDTA should not be added.
• The protease and phosphatase inhibitors in this product are harmful to humans. Handle with care and use appropriate protective measures to avoid direct contact with the body or inhalation.
• This product is for scientific research use by professionals only. It is not for clinical diagnosis or treatment, not for use in food or drugs, and should not be stored in ordinary residences.
• For your safety and health, please wear a lab coat and disposable gloves when handling this product.

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

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