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SKU AR0101
Pack size 100 mL (100 assays for 0.1g of tissuе)
Physical State Liquid
Applications Immunoassays: WB, ELISA, RIA, immunoprecipitation; reporter assays; protein kinase assays; protein purifications; folding studies; chromatographic studies; DNA-protein interaction assays

Overview

Product Name Mammalian Tissue Protein Extraction Reagent
SKU/Catalog Number AR0101
Physical State Liquid
Pack Size 100 mL (100 assays for 0.1g of tissuе)
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

Description

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.

Background

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)

• 2 ml 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.

Precautions

•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.

Mammalian Tissue Protein Extraction Reagent Images

Click the images to enlarge.

Mammalian Tissue Protein Extraction Reagent
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.
Mammalian Tissue Protein Extraction Reagent
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).
Mammalian Tissue Protein Extraction Reagent
Mammal Tissue Protein Extraction Reagent (AR0101)
100 mL per pack, Solution (liquid)
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Publications

The function of Notch1 intracellular domain in the differentiation of gastric cancer
An Aza resveratrol?chalcone derivative 6b protects mice against diabetic cardiomyopathy by alleviating inflammation and oxidative stress
HIF-1? contributes to hypoxia adaptation of the naked mole rat
Upregulation of Cdh1 Attenuates Isoflurane-Induced Neuronal Apoptosis and Long-Term Cognitive Impairments in Developing Rats
Estradiol-enhanced osteogenesis of rat bone marrow stromal cells is associated with the JNK pathway
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[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
MicroRNA-27a-3p Inhibits Melanogenesis in Mouse Skin Melanocytes by Targeting Wnt3a
The Protective Effects of Shen-Fu Injection on Experimental Acute Pancreatitis in a Rat Model
Expression patterns of?prdm1?during chicken embryonic and germline development
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 mole rat
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 Impairments in Developing Rats
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
Jin, G., Guo, L., Zhang, Y., Xue, Y., Zhang, X., Wang, X.,…, & Cheng, J. (2017). Expression and localization of lipocalin-type-prostaglandin D synthase in the goat testis, epididymis and sperm. Small Ruminant Research, 154, 1-4. Advance online publication. doi: 10.1016/j.smallrumres.2017.06.020
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 growth factor-regulated fibroblast migration.
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-JNK-Bcl-2 Pathway.
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.
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 ?? Protein...
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.

Customer Q&As

Q: Can AR0101 be used with preserved tissues?
A: AR0101 is not suitable for use on preserved tissues. We suggest you sonicate and freeze-thaw your samples to obtain proteins.