Altman:Protocols/Preparing Reagents/Making CPK: Difference between revisions
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David Altman (talk | contribs) (New page: {{Template:Altman}} <div style="padding: 10px; width: 700px; border: 5px solid #B22222;"> ==Making Creatine Phosphokinase (CPK)== *Creatine Phosphokinase – Calbiochem, Cat #: 238395, ...) |
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1. Make ABSA + 50% GlyOH | 1. Make ABSA + 50% GlyOH | ||
<table id="Table 1" border="1" cellpadding="3" cellspacing="0"> | |||
10x AB stock 500 | |||
10x BSA stock 500 | <tr> | ||
100x DTT stock 50 | <th><tt> </tt></th> | ||
GlyOH 2.5 mL | <th><tt>5 mL ABSA + GlyOH </tt></th> | ||
ddwater | </tr> | ||
<tr> | |||
<th><tt> 10x AB stock </tt></th> | |||
<th><tt> 500 uL </tt></th> | |||
</tr> | |||
<tr> | |||
<th><tt> 10x BSA stock </tt></th> | |||
<th><tt> 500 uL </tt></th> | |||
</tr> | |||
<tr> | |||
<th><tt> 100x DTT stock </tt></th> | |||
<th><tt> 50 uL </tt></th> | |||
</tr> | |||
<tr> | |||
<th><tt> GlyOH </tt></th> | |||
<th><tt> 2.5 mL </tt></th> | |||
</tr> | |||
<tr> | |||
<th><tt> ddwater </tt></th> | |||
<th><tt> fill to 5 mL </tt></th> | |||
</tr> | |||
</table> | |||
2. Chill the ABSA + GlyOH at -20°C | 2. Chill the ABSA + GlyOH at -20°C | ||
3. Add 1.5 mL of ABSA + GlyOH to CPK container and mix | 3. Add 1.5 mL of ABSA + GlyOH to CPK container and mix | ||
4. Aliquot 50 μL stocks | |||
4. Aliquot 50 μL stocks on ice | |||
5. Store at -20°C | 5. Store at -20°C | ||
Revision as of 17:56, 18 November 2012
Department of Physics, Willamette University
Making Creatine Phosphokinase (CPK)
- Creatine Phosphokinase – Calbiochem, Cat #: 238395, 5KU, store at -20°C
- Final stock (100x) should be 10 mg/mL stored in 0.1% BSA, 50% GlyOH, and AB conditions.
1. Make ABSA + 50% GlyOH
| 5 mL ABSA + GlyOH | |
|---|---|
| 10x AB stock | 500 uL |
| 10x BSA stock | 500 uL |
| 100x DTT stock | 50 uL |
| GlyOH | 2.5 mL |
| ddwater | fill to 5 mL |
2. Chill the ABSA + GlyOH at -20°C
3. Add 1.5 mL of ABSA + GlyOH to CPK container and mix
4. Aliquot 50 μL stocks on ice
5. Store at -20°C
