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Proteoloc™ Protease Inhibitor Cocktail EDTA Free

Protection From Endogenous Proteases During Protein Extraction

The process of breaking open cell walls and subcellular compartments during protein extraction liberates endogenous proteases. Without effective inhibition, these enzymes will quickly diminish yields of intact proteins from cell and tissue extracts. The addition of protease inhibitors to lysis buffers used for protein extractions is an effective means of protecting extracted proteins from enzymatic degradation by endogenous proteases.

Use Proteoloc™ EDTA free if the next step in your workflow is isoelectric focusing, metal affinity column separation, or another technique that is incompatible with chelation. Otherwise, use the standard Proteoloc™ Protease Inhibitor Cocktail, which contains EDTA. Both products offer protection against loss of intact proteins due to endogenous protease activity. Proteoloc™ Protease Inhibitor Cocktails are provided as 100X stock solutions in DMSO.

Protease Inhibitor Cocktail Proteloc Contains the Following Protease Inhibitors:

  • AEBSF: Irreversible serine protease inhibitor. Inhibits chymotrypsin, kallikrein, plasmin, thrombin, and trypsin
  • Aprotinin: Reversible serine protease inhibitor. Inhibits chymotrypsin, kallikrein, plasmin, trypsinogen, and urokinase
  • Bestatin: Metalloprotease inhibitor selective for aminopeptidases. Inhibits activity of leucine aminopeptidase, aminopeptidase B, and triamino peptidase
  • E-64: Irreversible inhibitor of cystine proteases. Inhibits calpain, papain, and cathespin B
  • Leupeptin: Reversible inhibitor of serine and cystine proteases. Inhibits calpain, papain, trypsin, and cathespin B
  • Pepstatin A: Reversible aspartic acid protease inhibitor. Inhibits pepsin, rennin, cathespin D, chymosin, and protease B.

Proteoloc™ Inhibitor Cocktail is the protease inhibitor mixture recommended for use with UPX Universal Protein Extraction Kits.

What Does the Proteoloc Protease Inhibitor Cocktail Do?

The Proteoloc Protease Inhibitor Cocktail protects proteins from endogenous protease activity during extraction.

Is the Proteoloc Protease Inhibitor Cocktail Compatible with My
Sample Cells and Tissues?

Yes. The Proteoloc Protease Inhibitor Cocktail is suitable for use during protein extraction from mammalian tissues and cells, yeasts, fungal cells, and bacteria.

What are the Compositions of the Proteoloc Protease Inhibitor Cocktail?

The composition of the Proteoloc Protease Inhibitor Cocktail is:
• AEBSF: Irreversible serine protease inhibitor. Inhibits chymotrypsin, kallikrein, plasmin, thrombin, and trypsin.
• Aprotinin: Reversible serine protease inhibitor. Inhibits chymotrypsin,
kallikrein, plasmin, trypsinogen, and urokinase.
• Bestatin: Metalloprotease inhibitor selective for aminopeptidases.
Inhibits activity of leucine aminopeptidase, aminopeptidase B, and triamino peptidase.
• EDTA: Inhibits metalloproteases by chelating divalent metal ions.
• E-64: Irreversible inhibitor of cystine proteases. Inhibits calpain, papain, and cathespin B.
• Leupeptin: Reversible inhibitor of serine and cystine proteases. Inhibits calpain, papain, trypsin, and cathespin B.
• Pepstatin A: Reversible aspartic acid protease inhibitor. Inhibits pepsin, rennin, cathespin D, chymosin, and protease B.
• DMSO

Note: The composition of the Proteoloc Protease Inhibitor Cocktail EDTA-free is the same as above, but without EDTA.

Which Proteoloc Protease Inhibitor Cocktail Should I Use?

Two protease inhibitor products are available from Protein Discovery: Proteoloc Protease Inhibitor Cocktail, and Proteoloc Protease Inhibitor Cocktail EDTA-free. Use the EDTA-free product if your workflow includes techniques that will not work in the presence of EDTA, such as isoelectric focusing or metal-affinity column separations. Otherwise, use Proteoloc Protease Inhibitor Cocktails the Proteoloc product with EDTA. For protein extractions intended for fractionation using the Gelfree 8100 Fractionation System, use Proteoloc Protease Inhibitor Cocktail with EDTA.

How Much Proteoloc Protease Inhibitor Cocktail Do I Need?

Proteoloc Protease Inhibitor Cocktails are supplied as 100X stock solutions, and should be diluted into the working volume of the lysis buffer you’re using for extraction. For example, if your next experiment involves extracting proteins from six tissue samples using 10 mL lysis buffer per sample, you will need 600 μL of Proteoloc Protease Inhibitor Cocktail.

What is Shelf Life of Proteoloc Protease Inhibitor Cocktails?

The shelf life of Proteoloc Protease Inhibitor Cocktails is eight months when stored at 4 °C, and two years when stored at -20 °C.

What Are the Advantages of Single-Use Proteoloc Packaging?

Single-use packaging helps you to complete your protein extraction experiments more quickly by providing Proteoloc Protease Inhibitor Cocktails in a pre-measured quantity container that speeds thawing and eliminates the need to measure out a 100 μL volume of a DMSO solvent solution.

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