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T-PER Tissue Protein Extraction Reagent  |
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An extra-mild tissue cell lysis solution designed for total protein extraction from tissue samples.
Biological tissue is a series of interconnected cells that perform a similar function within an orgnanism. To further analyze the function of the tissue and each cell within, it is necessary to lyse the cells and study the individual proteins. This can be difficult as tissue is quite dense and especially with interconnections between cells. Methods of tissue and cellular disruption require the use of physical means such as mortar and pestle, shearing, sonication and processors. Tissue can also be manipulated by temperature or freeze/thawing which causes the cell adhesion properties to weaken and allow for cell lysis into buffer. Physical disruption of tissue at cold temperatures (0°C - 4°C) helps to slow down protein degradation and keep some protein interactions intact. Regardless of the method chosen for cellular disruption, each method can be easily facilitated using the T-PER Tissue Protein Extraction Reagent.
This reagent utilizes a proprietary detergent in 25 mM bicine, 150 mM sodium chloride (pH 7.6) for tissue cell lysis. The simple composition of this reagent provides versatility for many different applications. Depending on the application, it may be advantageous to add other components such as protease inhibitors, salts, reducing agents, chelating agents, etc., to the reagent before proceeding with the cell lysis. The cell lysate prepared with this reagent may be used for reporter assays (e.g., luciferase, b -galactosidase, chloramphenicol acetyl transferase), protein kinase assays (e.g., PKA, PKC, tyrosine kinase), immunoassays (e.g., Western blots, ELISAs, RIAs), protein purifications and protein microarrays.
Highlights
- Simple procedure – just homogenize tissue sample in 1:20 (w/v) of tissue to T-PER Reagent, then centrifuge to pellet cell/tissue debris
- Mild detergent is dialyzable for quick and easy removal
- Can be used with additional components (e.g., protease inhibitors, salts, reducing agents, chelating agents, etc.)
- Lysate may be used for reporter assays, protein kinase assays, immunoassays and/or protein purifications
- The lysate is compatible with standard protein assays such as Pierce Coomassie Plus – The Better Bradford Protein Assay (Product # 23236).
References
- Aldred, M.A., et al. (2003). Cancer Res. 63, 2864-2871.
- Chakravarti, S. et al. (2003). Invest. Ophth. & Vis. Sci., 44(6), 2422-2432.
- Jepsen, K.H., et al. (2002). J. Biol. Chem. 277, 35532-35540.
- Lukashevich, I.S., et al. (2003). J. Virol. 77(3), 1727-1737.
- Paweletz, C., et al. (2001). Oncogene 20, 1981 - 1989.
- Runkuan, Y., et al. (2002). Am J Physiol Gastrointest Liver Physiol 283, G212–G221
- Runkuan, Y., et al. (2002). Amer. J. Pathol. 161, 1475-1484.
- Sheng, J.G., et al. (2002). J. Neurosci. 22(22), 9794-9799.
- Smallwood, A, et al. (2003). Biol. of Reprod. 69, 286–293.
- Yan, W. et al. (2003). PNAS 100(12), 7189-7194.
- Zen, Y. et al. (2002). American Journal of Pathology 161(4), 1475 - 1484,
Related Products
Mammalian Protein Extraction Reagent
Nuclear and Cytoplasmic Extraction Reagents
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Eukaryotic Membrane Protein Extraction Kit
Mitochondria Isolation Kit for Cultured Cells
Halt Protease Inhibitor Cocktail
Related Links
Poppers Cell Lysis Handbook
Remove detergent from protein samples
Cell Lysis Technical Information
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