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Product Info Summary
|Pack size||100 mL (100 assays for 0.1g of tissue)|
|Product Name||Mammalian Tissue Protein Extraction Reagent|
|Pack Size||100 mL (100 assays for 0.1g of tissue)|
|Recommended working concentration||1mL of extraction reagent per 0.1g of tissuе for tissue samples homogenization|
|Storage & Expiration||Upon receipt store at 4°C. Mammalian Tissue Protein Extraction Reagent is stable for one year. Product is shipped on ice.|
|Equivalent||Thermofisher (Product No. 78510); Millipore Sigma (Product No. C3228)|
|Description||Boster’s Mammalian Tissue Protein Extraction Reagent is a ready-to-use Western blot related reagent solution used for efficient extraction of total soluble protein in nondenatured state from animal tissues.|
|Application||Immunoassays: WB, ELISA, RIA, immunoprecipitation; reporter assays; protein kinase assays; protein purifications; folding studies; chromatographic studies; DNA-protein interaction assays
*Our Boster Guarantee covers the use of this product in the above tested applications.
|Cite This Product||Mammalian Tissue Protein Extraction Reagent (Boster Biological Technology, Pleasanton CA, USA, Catalog # AR0101)|
|Reagent Type||Universal tissue extraction buffer, detergent solution, cell lysis buffer|
|Usage||Obtaining homogenous tissue cell lysates with soluble protein extracts|
|Reactivity/severeness||Bland, minimal amount of detergent, detergent is dialyzable|
|Enzymatic Activity||None; use additives to add function|
|Additional components compatibility||Compatible with additives, e.g. protease inhibitors (PMSF), reducing agents, chelating agents, salts|
Mammalian Tissue Protein Extraction Reagent is a ready-to-use Western blot related reagent solution used for efficient extraction of total soluble protein in nondenatured state from animal tissues by homogenization and cell lysis utilizing a dialyzable mild detergent at low concentration, for maximal protein solubilization and minimal interference with protein biological activity enabling direct use in downstream analysis.
Extraction of tissue proteins being an essential step in the process of protein sample preparation is critical to successful protein analysis. Successful tissue protein extraction requires efficient cell lysis and protein solubilization, while preserving protein integrity and immunoreactivity. The varying tissue composition in different tissue samples implies different approaches for efficient and homogenous tissue protein extraction. The Mammalian Tissue Protein Extraction Reagent was designed for use with all tissues, independent of mechanical extraction methods. A mild nondenaturing and easily dialyzable detergent is added to the solution that aids in dissolving cell membranes and enhances solubilization of total protein without compromising function, and the neutral pH and salt concentration of the reagent enable more efficient extraction from cellular compartments yielding homogeneous lysates. The produced cell lysates are directly compatible with reporter gene expression assays (luciferase, beta-galactosidase, chloramphenicol acetyl transferase, CAT, alkaline phosphatase), protein kinase assays (PKA, PKC, tyrosine kinase), phosphatase assays (general phosphatases, tyrosine phosphatases), immunoassays ( Western blots, ELISAs, RIAs, immunoprecipitation), Coomassie-Blue and silver staining, protein purification procedures, electrophoresis, folding studies, chromatographic studies, DNA-protein interaction assays (gel-shift assays), and many other downstream applications.
Additional Materials Required
• Protease inhibitor (Product No. AR1182) and phosphatase inhibitor (Product No. AR1183)
• 2mL microcentrifuge tubes
• Tissue homogenizer
• Microcentrifuge capable of spinning at 10,000 x g
Procedure for Tissue Lysis
Note: Pre-chill an appropriate volume of Mammalian Tissue Protein Extraction Reagent at 4°C. If desired, add protease inhibitor and phosphatase inhibitor to the lysis buffer immediately before use.
1. Place the fresh tissue into chilled PBS and rinse several times. Mince the tissue into small pieces.
2. Add Mammalian Tissue Protein Extraction Reagent to the tissue at 10:1. (i.e., Add 10 mL chilled lysis buffer per gram of tissue.) Use a smaller volume of reagent if a more concentrated protein extract is required.
3. Homogenize for several minutes at high speed until no tissue chunks remain.
4. Incubate on ice for 30 minutes.
5. Centrifuge at ~10000 x g for 10 minutes.
6. Transfer supernatant to a new tube for further analysis.
• All steps of protein lysis should be operated on ice or at 4°C.
• Use BCA Protein Assay kit (Product No. AR0146) to quantify lysed proteins. Bradford Protein Assay kit is not recommended.
Related Boster Products
AR1182 Broad Spectrum Protease Inhibitor Cocktail
AR1183 Broad Spectrum Phosphatase Inhibitor Cocktail
AR0146 BCA Protein Assay Kit
AR0138 SDS-PAGE Gel Preparation Kit
AR1170 Hypersensitive ECL Chemiluminiscence Substrate
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Figure 1. Tissue cell lysis protein yield with Mammalian Tissue Protein Extraction Reagent
Mouse tissue proteins were extracted from different mouse tissues following the Mammalian Tissue Protein Extraction Reagent protocol. The protein concentration of each lysate was determined by BCA Protein Assay Kit (Product No. AR0146) to determine protein yield per milligram of starting tissue.
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Figure 2. Protein extraction from mouse tissues using Mammalian Tissue Protein Extraction Reagent
Proteins were extracted from mouse tissues following the Mammalian Tissue Protein Extraction Reagent protocol. Lysates (20ug) were separated by SDS-PAGE and transferred to a nitrocellulose membrane. Incubate with primary antibodies. Images were generated using Boster Hypersensitive ECL chemiluminiscence substrate (Product No. AR1170).
AR0101 has been cited in 26 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.
Tu Y,Chen D,Pan T,Chen Z,Xu J,Jin L,Sheng L,Jin X,Wang X,Lan X,Ge Y,Sun H,Chen Y.Inhibition of miR-431-5p attenuated liver apoptosis through KLF15/p53 signal pathway in S100 induced autoimmune hepatitis mice.Life Sci.2021 Jun 7:119698.doi:10.1016/j.lfs.2021.119698.Epub ahead of print.PMID:34111466.
AR0101 usage in article: APP:WB, SAMPLE:LIVER TISSUE, DILUTION:NA
Liu,B.,Dong,J.,Wang,S.,Yu,H.,Li,Z.,Sun,P.,& Zhao,L.(2021).Helicobacter pylori causes delayed gastric emptying by decreasing interstitial cells of Cajal.Experimental and Therapeutic Medicine,22,663.https://doi.org/10.3892/etm.2021.10095
AR0101 usage in article: APP:WB, SAMPLE:GASTRIC 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.
AR0101 usage in article: APP:WB, SAMPLE:HIPPOCAMPUS, DILUTION:NA
Li W,Yang C,Mei X,Huang R,Zhang S,Tang Y,Dong Q,Zhou C.Effect of the polyphenol-rich extract from Allium cepa on hyperlipidemic sprague-dawley rats. J Food Biochem.2020 Nov 20:e13565.doi:10.1111/jfbc.13565. Epub ahead of print.PMID:33219537.
Species: Rat,Allium cepa
AR0101 usage in article: APP:WB, SAMPLE:LIVER, DILUTION:NA
Xu J, Chen D, Jin L, Chen Z, Tu Y, Huang X, Xue F, Xu J, Chen M, Wang X, Chen Y. Ubiquitously specific protease 4 inhibitor-Vialinin A attenuates inflammation and fibrosis in S100-induced hepatitis mice through Rheb/mTOR signalling. J Cell Mol Med. 2020 D
Species: Mouse, Human
AR0101 usage in article: APP:WB, SAMPLE:AML12 CELLS, DILUTION:NA
Estradiol-enhanced osteogenesis of rat bone marrow stromal cells is associated with the JNK pathway
Decorin reduces hypertrophic scarring through inhibition of the TGF-?1/Smad signaling pathway in a rat osteomyelitis model
[Gly14]-Humanin Protects Against Amyloid ? Peptide-Induced Impairment of Spatial Learning and Memory in Rats
Polymorphisms in the CHIT1 gene: Associations with colorectal cancer
Molecular Cloning, mRNA Expression, and Localization of the G-protein Subunit Galphaq in Sheep Testis and Epididymis
Carnosine markedly ameliorates H9N2 swine influenza virus-induced acute lung injury
Xiao B, Wang S, Yang G, Sun X, Zhao S, Lin L, Cheng J, Yang W, Cong W, Sun W, Kan G, Cui S. Oncotarget. 2017 Nov 30;8(66):109941-109951. doi: 10.18632/oncotarget.22767. eCollection 2017 Dec 15. HIF-1α contributes to hypoxia adaptation of the naked...
Li X, Wei K, Hu R, Zhang B, Li L, Wan L, Zhang C, Yao W. Front Cell Neurosci. 2017 Nov 23;11:368. doi: 10.3389/fncel.2017.00368. eCollection 2017. Upregulation of Cdh1 Attenuates Isoflurane-Induced Neuronal Apoptosis and Long-Term Cognitive Impair...
Hu S, Chen Q, Lin T, Hong W, Wu W, Wu M, Du X, Jin R. Oncol Lett. 2018 May;15(5):6171-6178. doi: 10.3892/ol.2018.8118. Epub 2018 Feb 26. The function of Notch1 intracellular domain in the differentiation of gastric cancer
You S, Qian J, Sun C, Zhang H, Ye S, Chen T, Xu Z, Wang J, Huang W, Liang G. J Cell Mol Med. 2018 Mar;22(3):1931-1943. doi: 10.1111/jcmm.13477. Epub 2018 Jan 12. An Aza resveratrol
Zhu ZX, Sun CC, Ting Zhu Y, Wang Y, Wang T, Chi LS, Cai WH, Zheng JY, Zhou X, Cong WT, Li XK, Jin LT. Exp Cell Res. 2017 Mar 28. pii: S0014-4827(17)30181-7. doi: 10.1016/j.yexcr.2017.03.054. Hedgehog signaling contributes to basic fibroblast growt...
Zhu Z, Huang Y, Lv L, Tao Y, Shao M, Zhao C, Xue M, Sun J, Niu C, Wang Y, Kim S, Cong W, Mao W, Jin L. J Cell Physiol. 2017 Mar 28. doi: 10.1002/jcp.25934. Acute Ethanol Exposure-induced Autophagy-mediated Cardiac Injury via Activation of the ROS-...
Zhao Y, Wang P, Meng J, Ji Y, Xu D, Chen T, Fan R, Yu X, Yao J,, Dong C. Int J Mol Sci. 2015 May 14;16(5):10921-33. Doi: 10.3390/Ijms160510921. Microrna-27A-3P Inhibits Melanogenesis In Mouse Skin Melanocytes By Targeting Wnt3A.
Gui J, Xiong F, Yang W, Li J, Huang G. Am J Reprod Immunol. 2012 May;67(5):383-90. Doi: 10.1111/J.1600-0897.2011.01097.X. Epub 2012 Jan 9. Effects Of Acupuncture On Lif And Il-12 In Rats Of Implantation Failure.
Huang L, Cao J. Oxid Med Cell Longev. 2014;2014:248786. Doi: 10.1155/2014/248786. Epub 2014 Mar 5. The Protective Effects Of Shen-Fu Injection On Experimental Acute Pancreatitis In A Rat Model.
Gui J, Xiong F, Li J, Huang G. Evid Based Complement Alternat Med. 2012;2012:893023. Doi: 10.1155/2012/893023. Epub 2012 Jan 23. Effects Of Acupuncture On Th1, Th2 Cytokines In Rats Of Implantation Failure.
Chen Zy, Li J, Huang Gy. J Huazhong Univ Sci Technolog Med Sci. 2013 Apr;33(2):293-302. Doi: 10.1007/S11596-013-1114-Y. Epub 2013 Apr 17. Effect Of Bushen Yiqi Huoxue Recipe On Placental Vasculature In Pregnant Rats With Fetal Growth Restriction I...
Zhou J, Qu C, Sun Q, Wu L, Liu Y, Yang Z, Zhang J. Chem Biol Interact. 2014 Aug 5;219:57-63. Doi: 10.1016/J.Cbi.2014.05.009. Epub 2014 May 28. Sophoricoside Fails The Embryo Implantation By Compromising The Uterine Endometrial Receptivity At Impla...
Gao D, Ding F, Lei G, Luan G, Zhang S, Li K, Wang D, Zhang L, Dai D. Mol Med Rep. 2015 Apr;11(4):3009-14. Doi: 10.3892/Mmr.2014.3111. Epub 2014 Dec 18. Effects Of Focal Mild Hypothermia On Thrombin-Induced Brain Edema Formation And The Expression ...
Cai Hy, H??lscher C, Yue Xh, Zhang Sx, Wang Xh, Qiao F, Yang W, Qi Js. Neuroscience. 2014 Sep 26;277:6-13. Doi: 10.1016/J.Neuroscience.2014.02.022. Epub 2014 Feb 27. Lixisenatide Rescues Spatial Memory And Synaptic Plasticity From Amyloid ?? Prote...
Wan Z, Rui L, Li Z. Cell Tissue Res. 2014 May;356(2):341-56. Doi: 10.1007/S00441-014-1804-1. Epub 2014 Apr 2. Expression Patterns Of Prdm1 During Chicken Embryonic And Germline Development.
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