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(TM)Tj
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( Protocol)Tj
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[(I.)-635(Introduction)]TJ
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(Tissue-Specific Protein Concentrates are electrophoresis-ready solutions\
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(o)Tj
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(C, and protease inhibitors are present throughout the isolation procedur\
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(Each tube contains a solution of total protein in sample buffer consisti\
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(HCl, 2% SDS, 10% glycerol, 0.1% bromophenol blue \(pH 6.8\) \(1\). This \
protein solution)Tj
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(tion of your Protein Medley.)Tj
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[(II.)-600(Sample preparation for one dimensional SDS-polyacrylamide gel electropho\
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(sis \(PAGE\))Tj
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0.189 Tw 0.3 -1.4 Td
(The following procedure is based on the discontinuous polyacrylamide gel\
system)Tj
-0.041 Tw T*
(described by Laemmli \(1\). If an alternate electrophoresis system, or o\
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(employed, the sample preparation should be modified accordingly.)Tj
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[(A.)-590(General considerations)]TJ
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[(1.)-905(It is not advisable to repeatedly freeze and thaw the protein concentrat\
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(if small amounts of the concentrate will be used at a given time we reco\
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(C.)Tj
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[(3.)-905(Electrophoresis grade chemicals should be used for the preparation of al\
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[(4.)-905(The amount of total protein loaded per lane must be optimized for the pa\
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(m)Tj
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(tissue homogenate.)Tj
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(B. Additional materials required)Tj
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3.1 -1.4 Td
(Note: The following materials are required but not supplied.)Tj
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[(\245)-1195(Sample buffer \(pH 6.8\))]TJ
1.55 -1.4 Td
[(62.5 mM Tris)-11509(0.76)-1635(g)]TJ
0 -1.4 TD
[(2% \(w/v\) SDS)-11119(2.0)-2195(g)]TJ
0 Tw T*
[(10% \(v/v\) glycerol)-9364(10)-2480(ml)]TJ
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[(0.1% \(w/v\) bromophenol blue)-4262(0.1)-2195(g)]TJ
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(Add Tris, SDS, and glycerol to 75 ml of ddH)Tj
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(2)Tj
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(O.)Tj
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(Adjust pH to 6.8 with dilute HCl.)Tj
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(2)Tj
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(O. Store)Tj
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(buffer at room temperature.)Tj
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(2)Tj
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(O, mix gently until dissolved.)Tj
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(O.)Tj
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