Altman:Protocols/Preparing Reagents

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(New page: {{Template:Altman}} <div style="padding: 10px; width: 700px; border: 5px solid #B22222;"> =DESCRIPTION OF REAGENTS= Assay Buffer (AB) This contains the salts and the buffer that keep th...)
Current revision (13:18, 12 March 2013) (view source)
 
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=DESCRIPTION OF REAGENTS=
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='''''Preparing Reagents'''''=
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Assay Buffer (AB)
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=='''[[/Making assay buffer|Making assay buffer]]'''==
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This contains the salts and the buffer that keep the biological samples happy.
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Contents of AB: 25 mM KCl, 25 mM Imidazole-Cl (pH 7.4), 1 mM EGTA, 4 mM MgCl2, 10 mM DTT
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AB is made using a 10x AB stock and a 100x DTT stock
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Dithiothreitol (DTT)
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=='''[[/Making GOC|Making glucose oxidase catalase (GOC)]]'''==
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C4H10O2S2
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DTT is a strong reducing reagent; we use it to reduce the disulfide bonds of proteins and, more generally, to prevent intramolecular and intermolecular disulfide bonds from forming between cysteine residues of proteins
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=='''[[/Making GB|Making glucose + beta-mercaptoethanol (G/β)]]'''==
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DTT often crashes out of solution when it is frozen.  Take care to vortex the tube of DTT when you thaw an aliquot.
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=='''[[/Making CPK|Making creatine phosphokinase (CPK)]]'''==
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=='''[[/Making PCR|Making creatine phosphate disodium salt (PCr)]]'''==
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=='''[[/Making nitrocellulose coverslips|Making nitrocellulose beaded coverslips]]'''==
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=='''[[/Making ATP stock solution|Making 0.1 M ATP stock]]'''==
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Department of Physics, Willamette University

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Contents

Preparing Reagents

Making assay buffer

Making glucose oxidase catalase (GOC)

Making glucose + beta-mercaptoethanol (G/β)

Making creatine phosphokinase (CPK)

Making creatine phosphate disodium salt (PCr)

Making nitrocellulose beaded coverslips

Making 0.1 M ATP stock

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