|Pack size||1 mL|
|Applications|| Western blotting, protein purification, Co-IP; assays for protein expression, activity, modification, profiling and characterization, quantitative measurement; epitope tagging; reporter gene analysis|
|Product Name||Broad Spectrum Protease Inhibitor Cocktail|
|Synonyms||Broad Spectrum Protease Inhibitors|
|Description||Broad Spectrum Protease Inhibitor Cocktail (100X) is a Western blot related ready-to-use concentrated stock solution reagent containing a blend of seven protease inhibitors that is to be added to cell lysis buffer to protect the integrity and functionality of native cellular proteins against degradation by multiple classes of endogenous proteases during protein extraction and sample preparation procedures.|
|Application||Western blotting, protein purification, Co-IP; assays for protein expression, activity, modification, profiling and characterization, quantitative measurement; epitope tagging; reporter gene analysis|
|Pack Size||1 mL||Reagent Type||Western Blotting related reagent; Inhibitors||Usage||Protect native cellular proteins from destructive degradation by endogenous proteases following cell lysis
Preserve native cellular proteins intact and functional
Screen extracts for proteolytic activity
Study proteolysis in the regulation of cellular processes
|Content||E-64, AEBSF-HCL, bestatin, leupeptin, aprotinin, Pepstatin, and EDTA stabilized in dimethylsulfoxide (DMSO)||Target Specificity||Serine proteases, cysteine proteases, aspartic acid proteases, metalloproteases, aminopeptidases||Target Sample||Cell lysis extracts|
|Protease Inhibitor Component||MW||Protease Family Targeted||Inhibition Type||Typical Working (1X) Conc.||Concentration in 100X cocktail|
|E-64||357.4||Cysteine proteases (papain, calpain, lysosomal cathepsins)||Irreversible||1 to 20µM||1.5mM|
|AEBSF-HCI||239.5||Serine proteases (trypsin, chymotrypsin, plasmin, trypsinogen, urokinase, kallikrein)||Irreversible||0.2 to 1.0mM||100mM|
|Bestatin||308.4||Amino-peptidases||Reversible||1 to 10µM||5mM|
|Leupeptin||475.6||Serine and cysteine proteases||Reversible||10 to 100µM||2mM|
|Aprotinin||6511.5||Serine proteases (trypsin, chymotrypsin, plasmin, trypsinogen, urokinase, kallikrein)||Reversible||100 to 200nM||80µM|
|Pepstatin A||685.9||Aspartic acid proteases (pepsin and rennin)||Reversible||1 to 20µM||1mM|
|EDTA||372.2||Metalloproteases (thermolysin and carboxypeptidase A)||Reversible||2 to 10mM||2mM|
|Form Supplied||100X concentrated stock solution|
|Physical State||Liquid, colorless, transparent|
|Pack Size||1 ml|
|Recommended working concentration||100-fold dilution in tissue lysis buffer|
|10 µl of the Protease Inhibitor Cocktail solution is enough to inhibit degradation of proteins in 1 ml lysate|
|Compatibility with reagents||Fully compatible with cell lysis buffers and Broad Spectrum Phosphatase Inhibitor Cocktail|
|Compatibility with assays||Not MS-compatible: contain AEBSF;|
|Not compatible with immobilized metal chelate affinity chromatography and 2D gel electrophoresis: contain EDTA|
|Storage||Store at -20˚C for one year|
|Safety Precautions||Harmful. PMSF is toxic and causes irritation to the eyes and skin. Please operate with caution and wear eye and hand protection and proper lab garments. FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC AND CLINICAL USE|
Usage and Handling
The product is supplied as a 100X concentrated stock solution in a liquid format for improved accuracy, solubility, and ease of use in comparison to traditional tablets.
1. Add broad spectrum protease inhibitor cocktail in lysis buffer in the ratio of 1:100 and mix well.
2. Add the solution in to samples for protein extraction.
Since endogenous protease levels may vary among cell and tissue types, it is up to the researcher to optimize the concentration for their experiments.
Crude cell extracts contain a number of endogenous enzymes, such as proteases and phosphatases, which are capable of digesting the proteins present in the extract. An optimized method to improve the yield of intact and functional native proteins is to add inhibitors of these enzymes known to be present in the source material. This broad spectrum protease inhibitor cocktail is a complex of various protease inhibitors, which has been tested for inhibiting proteases and esterase broadly.
Protease inhibitors are critical reagents for the preservation of protein integrity during purification and analysis procedures by knocking out specific proteases to avoid peptide bond hydrolysis and subsequent protein destruction. They are biological or chemical compounds that function by reversibly or irreversibly binding to the protease. Most known proteases belong to one of four evolutionarily distinct enzyme families based on the functional groups involved in the peptide bond cleavage. No single chemical is effective for all types of proteases. A mixture or "cocktail" of several different inhibitor compounds is commonly used that consistently inhibit a multitude of protease classes to ensure that protein extracts do not degrade before analysis for target proteins of interest. Not using protease inhibitors leads to the loss of a large number of valuable proteins in a lysate sample, adversely affecting downstream applications by biologically meaningless representation of protein activities and gaining false negative immunostaining results for targets of interest.
Broad Spectrum Protease Inhibitor Cocktail (100x) Images
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