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Product Info Summary
List of Components
|Broad Spectrum Protease Inhibitor Cocktail||1||1mL (100X)||Containing AEBSF, aprotinin, bestatin, E-64, leupeptin and pepstatin A stabilized in dimethylsulfoxide (DMSO)||AR1182-A|
|0.1M EDTA solution||1||1mL (100X)||EDTA solution for optional metalloprotease inhibition||AR1182-B|
|Product Name||Broad Spectrum Protease Inhibitor Cocktail|
|Physical State||Liquid, colorless, transparent|
|Safety Precautions||Harmful. DMSO is toxic and causes irritation to the eyes and skin. Please operate with caution and wear eye and hand protection and proper lab garments.|
|Recommended working concentration||100-fold dilution in lysis buffer
10 μL of the Protease Inhibitor Cocktail solution is enough to inhibit degradation of proteins in 1 mL lysate.
|Storage||Upon receipt store at -20°C. It is stable for one year. Product is shipped on ice.|
|Equivalent||Thermofisher (Product No.78430, A32963)|
|Type of DAPI||Content||Catalog Number|
|Broad Spectrum Protease Inhibitor Cocktail||Broad Spectrum Protease Inhibitor Cocktail, 1mL (100X)|
0.1M EDTA solution, 1mL (100X)
|Broad Spectrum Protease Inhibitor Cocktail (EDTA free)||Broad Spectrum Protease Inhibitor Cocktail, 1mL (100X)||AR1182-1|
|Protease Inhibitor Component||MW||Protease Family Targeted||Inhibition Type||Typical Working (1X) Conc.|
|E-64||357.4||Cysteine proteases (papain, calpain, lysosomal cathepsins)||Irreversible||15μM|
|AEBSF||239.5||Serine proteases (trypsin, chymotrypsin, plasmin, trypsinogen, urokinase, kallikrein)||Irreversible||1mM|
|Leupeptin||475.6||Serine and cysteine proteases||Reversible||20μM|
|Aprotinin||6511.5||Serine proteases (trypsin, chymotrypsin, plasmin, trypsinogen, urokinase, kallikrein)||Reversible||800nM|
|Pepstatin A||685.9||Aspartic acid proteases (pepsin and rennin)||Reversible||10μM|
|EDTA||372.2||Metalloproteases (thermolysin and carboxypeptidase A)||Reversible||1mM|
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. 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.
|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
|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
|Target Specificity||Serine proteases, cysteine proteases, aspartic acid proteases, metalloproteases, aminopeptidases|
|Target Sample||Cell lysis extracts|
|Description||Boster’s Broad Spectrum Protease Inhibitor Cocktail is a complex of various protease inhibitors, which has been tested for inhibiting proteases and esterase broadly.|
|Cite This Product||Broad Spectrum Protease Inhibitor Cocktail (Boster Biological Technology, Pleasanton CA, USA, Catalog # AR1182)|
|Application|| Western blotting, protein purification, Co-IP; assays for protein expression, activity, modification, profiling and characterization, quantitative measurement; epitope tagging; reporter gene analysis |
*Our Boster Guarantee covers the use of this product in the above tested applications.
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.
Usage and Handling
• Equilibrate the bottle to room temperature before use.
• Vortex the bottle before use to ensure a homogeneous suspension
• This protease inhibitor cocktail is supplied at a 100X concentration in DMSO and is generally effective when used at a 1X final concentration; however, if a sample contains particularly high levels of proteases, the effective cocktail concentration might require optimization.
1. Add broad spectrum protease inhibitor cocktail in lysis buffer at a ratio of 1:100 and mix well.
2. Optional: For inhibition of metalloproteases, add 0.1M EDTA solution in lysis buffer at a ratio of 1:100 and mix well.
3. Add the solution in to cell or tissue samples for protein extraction.
Lysis buffer containing protease inhibitor cocktail should be freshly prepared before use.
Validation Images & Assay Conditions
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Protease activity for extracted protein from mouse tissue while using different protease inhibitor cocktails
AR1182 has been cited in 10 publications:
*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.
Apolipoprotein E4 Impairs in vivo Hippocampal Long-Term Synaptic Plasticity by Reducing the Phosphorylation of CaMKIIα and CREB
Overendocytosis of superparamagnetic iron oxide particles increases apoptosis and triggers autophagic cell death in human osteosarcoma cell under a spinning magnetic field
The hemagglutinin-neuramidinase protein of Newcastle disease virus upregulates expression of the TRAIL gene in murine natural killer cells through the activation of Syk and NF-κB
Functional Role of NRF2 in Cervical Carcinogenesis
Spinal 5-HT3 receptor contributes to somatic hyperalgesia induced by sub-chronic stress:
UNC5B Promotes Vascular Endothelial Cell Senescence via the ROS-Mediated P53 Pathway
SFKs/p38 Pathway is Involved in Radicular Pain by Promoting Spinal Expression of Pro-Inflammatory Cytokines in a Rat Model of Lumbar Disc Herniation
Capsaicin enhances the antitumor activity of sorafenib in hepatocellular carcinoma cells and mouse xenograft tumors through increased ERK signaling
Puerarin alleviate radicular pain from lumbar disc herniation by inhibiting ERK-dependent spinal microglia activation
Hydrogen sulfide reduces pyroptosis and alleviates ischemia-reperfusion acute kidney injury by inhibiting NLRP3 inflammasome
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