Altman:Protocols/Preparing Reagents/Making assay buffer: Difference between revisions

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==Making Assay Buffer==
Prepare the following beforehand:
*100x DTT stock: 100 mM dithiothreitol
*10x BSA stock: 10 mg/mL bovine serum albumin
*10x AB stock: 250 mM KCl, 250 mM Imidazole-Cl (pH 7.4), 10 mM EGTA, 40 mM MgCl<sub>2</sub>


<table id="Table 1" border="1" cellpadding="3" cellspacing="0">
<table id="Table 1" border="1" cellpadding="3" cellspacing="0">
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<tr>
<tr>
<th><tt> </tt></th>
<th><tt> </tt></th>
<th><tt>1 mL Assay Buffer</tt></th>
<th><tt>1 mL Assay Buffer
<th><tt>1 mL Assay Buffer + BSA</tt></th>
 
(AB)
</tt></th>
<th><tt>1 mL Assay Buffer + BSA
 
(ABSA)
</tt></th>
</tr>
</tr>


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<th><tt> 100 uL </tt></th>
<th><tt> 100 uL </tt></th>
<th><tt> 100 uL </tt></th>
<th><tt> 100 uL </tt></th>
</tr>
<tr>
<th><tt> 100x DTT stock </tt></th>
<th><tt> 10 uL </tt></th>
<th><tt> 10 uL </tt></th>
</tr>
<tr>
<th><tt> 10x BSA stock </tt></th>
<th><tt> -- </tt></th>
<th><tt> 100 uL </tt></th>
</tr>
<tr>
<th><tt> ddwater </tt></th>
<th><tt> 890 uL </tt></th>
<th><tt> 790 uL </tt></th>
</tr>
</tr>


</table>
</table>

Latest revision as of 18:43, 10 November 2012


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Making Assay Buffer

Prepare the following beforehand:

  • 100x DTT stock: 100 mM dithiothreitol
  • 10x BSA stock: 10 mg/mL bovine serum albumin
  • 10x AB stock: 250 mM KCl, 250 mM Imidazole-Cl (pH 7.4), 10 mM EGTA, 40 mM MgCl2


1 mL Assay Buffer

(AB)

1 mL Assay Buffer + BSA

(ABSA)

10x AB stock 100 uL 100 uL
100x DTT stock 10 uL 10 uL
10x BSA stock -- 100 uL
ddwater 890 uL 790 uL
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