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Product Info Summary
|Applications|| Western blotting, protein purification, Co-IP; assays for protein expression, activity, modification, profiling and characterization, quantitative measurement; epitope tagging; reporter gene analysis|
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.
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Protease activity for extracted protein from mouse tissue while using different protease inhibitor cocktails
AR1182 has been cited in 44 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.
The lncRNA CRNDE promotes colorectal cancer cell proliferation and chemoresistance via miR-181a-5p-mediated regulation of Wnt/β-catenin signaling
The activation and function of IL-36γ in neutrophilic inflammation in chronic rhinosinusitis
Yating Li,Jiongjie Jing,Wenqing Dang,Qi Han,Xiangyu Guo,Kaiqi Jia,Ying Cheng,Kai Wang,Ermias Kebreab,Lihua Lyu,Effects of Notch2 on proliferation, apoptosis and steroidogenesis in bovine luteinized granulosa cells,Theriogenology,Volume 171,2021,Pages 55-63,ISSN 0093-691X,https://doi.org/10.1016/j.theriogenology.2021.05.009.
Species: Holstein cow
AR1182 usage in article: APP:WB, SAMPLE:LGCS, DILUTION:NA
Jia Cui,Guangxin Chen,Zhonghua Zhao,Liqing Wang,Da Wen,Li Xing,Changxin Wu,ASAP1 regulates the uptake of Mycobacterium tuberculosis H37Ra in THP1-derived macrophages by remodeling actin cytoskeleton,Tuberculosis,Volume 129,2021,102090,ISSN 1472-9792,https://doi.org/10.1016/j.tube.2021.102090.
AR1182 usage in article: APP:WB, SAMPLE:THP-1 CELL, DILUTION:NA
Qian L,Yang X,Li S,Zhao H,Gao Y,Zhao S,Lv X,Zhang X,Li L,Zhai L,Zhou F,Chen B.Reduced O-GlcNAcylation of SNAP-23 promotes cisplatin resistance by inducing exosome secretion in ovarian cancer.Cell Death Discov.2021 May 18;7(1):112.doi:10.1038/s41420-021-00489-x.PMID:34001861.
AR1182 usage in article: APP:WB, SAMPLE:A2780 CELL AND SKOV3 CELL, DILUTION:NA
Mahmoud MF,Nabil M,Abdo W,Abdelfattah MAO,El-Shazly AM,El Kharrassi Y,Sobeh M.Syzygium samarangense leaf extract mitigates indomethacin-induced gastropathy via the NF-κB signaling pathway in rats.Biomed Pharmacother.2021 May 6;139:111675.doi:10.1016/j.biopha.2021.111675.Epub ahead of print.PMID:33965725.
AR1182 usage in article: APP:WB, SAMPLE:GASTRIC TISSUE, DILUTION:NA
Li Y, Zhang J, Shi J, Liu K, Wang X, Jia Y, He T, Shen K, Wang Y, Liu J, Zhang W, Wang T, Zheng Z, Hu D. Exosomes derived from human adipose mesenchymal stem cells attenuate hypertrophic scar fibrosis by miR-192-5p/IL-17RA/Smad axis. Stem Cell Res Ther. 2021 Mar 31;12(1):221. doi: 10.1186/s13287-021-02290-0. PMID: 33789737.
AR1182 usage in article: APP:WB, SAMPLE:FIBROBLASTS AND SKIN TISSUE, DILUTION:NA
Guo L, Li H, Fan T, Ma Y, Wang L. Synergistic efficacy of curcumin and anti-programmed cell death-1 in hepatocellular carcinoma. Life Sci. 2021 Mar 19:119359. doi: 10.1016/j.lfs.2021.119359. Epub ahead of print. PMID: 33753114.
AR1182 usage in article: APP:WB, SAMPLE:PBMCS, DILUTION:NA
Hu Y, Huang W, Luo Y, Xiang L, Wu J, Zhang Y, Zeng Y, Xu C, Meng X, Wang P. Assessment of the Anti-Inflammatory Effects of Three Rhubarb Anthraquinones in LPS-Stimulated RAW264.7 Macrophages Using a Pharmacodynamic Model and Evaluation of the Structure-Activity Relationships. J Ethnopharmacol. 2021 Mar 16: 114027.doi:10.1016/j.jep.2021.114027. Epub ahead of print. PMID: 33741438.
AR1182 usage in article: APP:WB, SAMPLE:RAW264.7 CELL, DILUTION:NA
Liu Hui,Zhou Chuankun,Qi Dahu,Gao Yutong,Zhu Meipeng,Tao Tenghui,Sun Xuying,Xiao Jun,Inhibiting Monoacylglycerol Lipase Suppresses RANKL-Induced Osteoclastogenesis and Alleviates Ovariectomy-Induced Bone Loss.Frontiers in Cell and Developmental Biology,VOLUME=9,YEAR=2021, PAGES=544, URL=https://www.frontiersin.org/article/10.3389/fcell.2021.640867. DOI=10.3389/fcell.2021.640867,ISSN=2296-634X
AR1182 usage in article: APP:WB, SAMPLE:BONE MARROW MACROPHAGES, DILUTION:NA
Lei L,Guo Y,Lin J,Lin X,He S,Qin Z,Lin Q. Inhibition of endotoxin-induced acute lung injury in rats by bone marrow-derived mesenchymal stem cells: Role of Nrf2/HO-1 signal axis in inhibition of NLRP3 activation. Biochem Biophys Res Commun.2021 Mar 10;551:7-13.doi:10.1016/j.bbrc.2021.03.009.Epub ahead of print.PMID: 33713981.
AR1182 usage in article: APP:WB, SAMPLE:LUNG TISSUE, DILUTION:NA
Zhang F,Liu X,Li B,Li Z,Grad S,Chen D,Gao M,Liu S.The effect of hyaluronic acid on nucleus pulposus extracellular matrix production through hypoxia-inducible factor-1α transcriptional activation of CD44 under hypoxia.Eur Cell Mater.2021 Feb 3;41:142-152.d
AR1182 usage in article: APP:WB, SAMPLE:NPCS, DILUTION:NA
Wenpeng Cao,Zhirui Zeng,Zhiwei He et al.Hypoxia-Related Gene FUT11 Promoted Pancreatic Cancer Progression Via Maintaining The Stabilization of PDK1,27 January 2021,PREPRINT (Version 1) available at Research Square [https://doi.org/10.21203/rs.3.rs-152464/
AR1182 usage in article: APP:IP, SAMPLE:LIVER TISSUE, DILUTION:NA
Jia X,Chen H,Ren Y,Dejizhuoga,Gesangyuzhen,Gao N,Feng H,Huang W,Liao Y,Yu H.BAP1 antagonizes WWP1-mediated transcription factor KLF5 ubiquitination and inhibits autophagy to promote melanoma progression.Exp Cell Res.2021 Jan 28:112506.doi:10.1016/j.yexcr.
AR1182 usage in article: APP:WB, SAMPLE:HUMAN SKIN MELANOMA CELL, DILUTION:NA
Nabil M, El Raey MA, Abdo W, Abdelfattah MAO, El-Shazly AM, Sobeh M, Mahmoud MF. Gastro-Protective Effects of Albizia anthelmintica Leaf Extract on Indomethacin-Induced Gastric Ulcer in Wistar Rats: In Silico and In Vivo Studies. Antioxidants (Basel). 202
AR1182 usage in article: APP:PROTEIN QUANTIFICATION, SAMPLE:STOMACH TISSUE, DILUTION:NA
Yuan L,Zhang J,Guo JH,Holscher C,Yang JT,Wu MN,Wang ZJ,Cai HY,Han LN,Shi H,Han YF,Qi JS. DAla2-GIP-GLU-PAL Protects Against Cognitive Deficits and Pathology in APP/PS1 Mice by Inhibiting Neuroinflammation and Upregulating cAMP/PKA/CREB Signaling Pathways.
AR1182 usage in article: APP:WB, SAMPLE:HIPPOCAMPUS, DILUTION:NA
Gang-Zhu Xu,Yang Xue,Si-Qi Wei,Jia-Heng Li,Richard J. Traub,Mao-De Wang,Dong-Yuan Cao,Valproate reverses stress-induced somatic hyperalgesia and visceral hypersensitivity by up-regulating spinal 5-HT2C receptor expression in female rats,Neuropharmacology,
AR1182 usage in article: APP:WB, SAMPLE:SPINAL CORD TISSUE, DILUTION:NA
Li M,Xie X,Chen H,Xiong Q,Tong R,Peng C,Peng F.Aconitine induces cardiotoxicity through regulation of calcium signaling pathway in zebrafish embryos and in H9c2 cells.J Appl Toxicol.2020 Jun;40(6):780-793.doi:10.1002/jat.3943.Epub 2020 Jan 23.PMID:3197543
AR1182 usage in article: APP:WB, SAMPLE:H9C2 CELL, DILUTION:NA
Wang X,Tong Y,Zhang J,Khan N,Zhang K,Bai H,Zhang Q,Chen Y.Neuroinflammation changes with periodontal inflammation status during periodontitis in wild-type mice.Oral Dis.2020 Aug 20.doi:10.1111/odi.13618.Epub ahead of print.PMID:32815656.
AR1182 usage in article: APP:ELISA, SAMPLE:HIPPOCAMPI, DILUTION:NA
Xu L,Shen J,Dai S,Sun L,Chen X. Tetramethylpyrazine Attenuated Sevoflurane-Induced Neurotoxicity by Enhancing Autophagy through GPR50/CREB Pathway in SH-SY5Y Cells. Am J Chin Med.2020;48(4):945-966.doi: 10.1142/S0192415X20500457.Epub 2020 May 30.PMID:3247
AR1182 usage in article: APP:WB, SAMPLE:SH-SY5Y CELL, DILUTION:NA
Su Q,Li T,He PF,Lu XC,Yu Q,Gao QC,Wang ZJ,Wu MN,Yang D,Qi JS.Trichostatin A ameliorates Alzheimer‘s disease-related pathology and cognitive deficits by increasing albumin expression and Aβ clearance in APP/PS1 mice.Alzheimers Res Ther.2021 Jan 4;13(
AR1182 usage in article: APP:WB, SAMPLE:HIPPOCAMPUL TISSUE, DILUTION:NA
Cai P,Lu Z,Jiang T,Wang Z,Yang Y,Zheng L,Zhao J.Syndecan-4 involves in the pathogenesis of rheumatoid arthritis by regulating the inflammatory response and apoptosis of fibroblast-like synoviocytes.J Cell Phys iol.2020 Feb;235(2):1746-1758.doi:10.1002/jcp
AR1182 usage in article: APP:WB, SAMPLE:FLSS, DILUTION:NA
Sun K,Luo J,Jing X,Xiang W,Guo J,Yao X,Liang S,Guo F,Xu T.Hyperoside ameliorates the progression of osteoarthritis: An in vitro and in vivo study.Phytomedicine.2020 Oct 14;80:153387.doi:10.1016/j.phymed.2020. 153387.Epub ahead of print.PMID:33130473.
AR1182 usage in article: APP:WB, SAMPLE:CHONDROCYTES, DILUTION:NA
Li Y,Zhu H,Pan L,Zhang B,Che H. microRNA-103a-3p confers protection against lipopolysaccharide-induced sepsis and consequent multiple organ dysfunction syndrome by targeting HMGB1. Infect Genet Evol. 2020 Dec 14:104681.doi: 10.1016/j.meegid.2020.104681.Ep
Li Q, Li B, Lu CL, Wang JY, Gao M, Gao W. LncRNA LINC01857 promotes cell growth and diminishes apoptosis via PI3K/mTOR pathway and EMT process by regulating miR-141-3p/MAP4K4 axis in diffuse large B-cell lymphoma. Cancer Gene Ther. 2020 Dec 12. doi: 10.10
AR1182 usage in article: APP:WB, SAMPLE:HCC1395 CELL, DILUTION:NA
Jiang C, Xie N, Sun T, Ma W, Zhang B, Li W. Xanthohumol Inhibits TGF-β1-Induced Cardiac Fibroblasts Activation via Mediating PTEN/Akt/mTOR Signaling Pathway. Drug Des Devel Ther. 2020 Dec 7;14:5431-5439. doi: 10.2147/DDDT.S282206. PMID: 33324040; PMCID: P
AR1182 usage in article: APP:WB, SAMPLE:FRESH MYOCARDIUM, DILUTION:NA
Spinal 5-HT3 receptor contributes to somatic hyperalgesia induced by sub-chronic stress Zhuo-Lin Li,1,2 Yang Xue,1 Zhuo-Ying Tao,1 Wen-Zhi Du,1 Yue-Gui Jiang,2 and Dong-Yuan Cao1 Mol Pain. 2019; 15: 1744806919859723. Published online 2019 Jul 5. doi:
The lncRNA CRNDE promotes colorectal cancer cell proliferation and chemoresistance via miR-181a-5p-mediated regulation of Wnt/?-catenin signaling
Overendocytosis of superparamagnetic iron oxide particles increases apoptosis and triggers autophagic cell death in human osteosarcoma cell under a spinning magnetic field
miR-20b Inhibits T Cell Proliferation and Activation via NFAT Signaling Pathway in Thymoma-Associated Myasthenia Gravis
[Gly14]-Humanin Protects Against Amyloid ? Peptide-Induced Impairment of Spatial Learning and Memory in Rats
Protection Efficacy of the Extract of Ginkgo biloba against the Learning and Memory Damage of Rats under Repeated High Sustained +Gz Exposure
The role of noggin in regulation of high glucose-induced apoptosis and insulin secretion in INS-1 rat beta cells
Liu W, Cui L, Xu H, Zhu Z, Gao X. Appl Environ Microbiol. 2017 Oct 31;83(22). pii: e01619-17. doi: 10.1128/AEM.01619-17. Print 2017 Nov 15. Flexibility-Rigidity Coordination of the Dense Exopolysaccharide Matrix in Terrestrial Cyanobacteria Acclim...
Li CJ, Sun B, Fang QH, Ding M, Xing YZ, Chen LM, Yu DM. Front Endocrinol (Lausanne). 2017 Nov 27;8:326. doi: 10.3389/fendo.2017.00326. eCollection 2017. Saxagliptin Induces β-Cell Proliferation through Increasing Stromal Cell-Derived Factor-1α In ...
Guo J, Li W, Wu Y, Jing X, Huang J, Zhang J, Xiang W, Ren R, Lv Z, Xiao J, Guo F. Front Pharmacol. 2017 Oct 4;8:693. doi: 10.3389/fphar.2017.00693. eCollection 2017. Meclizine Prevents Ovariectomy-Induced Bone Loss and Inhibits Osteoclastogenesis ...
Peng Z, Wang Y, Fan J, Lin X, Liu C, Xu Y, Ji W, Yan C, Su C. Sci Rep. 2017 Jan 24;7:41254. doi: 10.1038/srep41254. Costunolide and dehydrocostuslactone combination treatment inhibit breast cancer by inducing cell cycle arrest and apoptosis throug...
Wei Z, Junhong G, Xiaoyuan N, Jie W, Zhaojun W, Meina W, Wei Y, Jun Z, Jinshun Q. Biomed Res Int. 2017;2017:1238687. doi: 10.1155/2017/1238687. Epub 2017 Sep 18. Citalopram Ameliorates Impairments in Spatial Memory and Synaptic Plasticity in Femal...
Liu D, Zhou Y, Peng Y, Su P, Li Z, Xu Q, Tu Y, Tian X, Yang H, Wu Z, Mei W, Gao F. Front Mol Neurosci. 2018 Mar 6;11:72. doi: 10.3389/fnmol.2018.00072. eCollection 2018. Endoplasmic Reticulum Stress in Spinal Cord Contributes to the Development of...
Liang, Y., Song, D.Z., Liang, S., Zhang, Z.F., Gao, L.X., & Fan, X.H. (2017). The hemagglutinin-neuramidinase protein of Newcastle disease virus upregulates expression of the TRAIL gene in murine natural killer cells through the activation of Syk ...
He Ql, Chen Y, Qin J, Mo Sl, Wei M, Zhang Jj, Li Mn, Zou Xn, Zhou Sf, Chen Xw, Sun Lb. Med Sci Monit. 2012 Jun;18(6):Br229-36. Osthole, A Herbal Compound, Alleviates Nucleus Pulposus-Evoked Nociceptive Responses Through The Suppression Of Overexpr...
Dong L, Wang X, Tan J, Li H, Qian W, Chen J, Chen Q, Wang J, Xu W, Tao C, Wang S. J Cell Mol Med. 2014 Nov;18(11):2213-24. Doi: 10.1111/Jcmm.12353. Epub 2014 Aug 28. Decreased Expression Of Microrna-21 Correlates With The Imbalance Of Th17 And Tre...
Wan J, Xia L, Liang W, Liu Y, Cai Q. J Diabetes Res. 2013;2013:647107. Doi: 10.1155/2013/647107. Epub 2013 Apr 9. Transplantation Of Bone Marrow-Derived Mesenchymal Stem Cells Promotes Delayed Wound Healing In Diabetic Rats.
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