User:Timothy L. Foley/Notebook/refolding matrix/2009/10/13: Difference between revisions
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DTT<BR> | DTT<BR> | ||
TCEP<BR> | TCEP<BR> | ||
none<BR> | none<BR> | ||
<BR> | <BR> | ||
<u>additives</u>:<BR> | <u>additives</u>:<BR> | ||
arginine/asparate/none<BR> | |||
arginine | |||
asparate<BR> | |||
divalent cations (2 Mg++/ 0.1 Ca++)<BR> | divalent cations (2 Mg++/ 0.1 Ca++)<BR> | ||
GnCl<BR> | GnCl<BR> | ||
Ligand<BR> | Ligand<BR> | ||
PEG 3350 | PEG 3350/Glycerol/Sucrose<BR> | ||
Glycerol | |||
Sucrose<BR> | |||
''BSA''<BR> | ''BSA''<BR> | ||
NaCl/KCl (high ionic strength)<BR> | NaCl/KCl (high ionic strength)<BR> | ||
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=== | ===salt mix=== | ||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''salt mix''' | |||
| align="center" style="background:#f0f0f0;"|'''''' | |||
| align="center" style="background:#f0f0f0;"|'''''' | |||
| align="center" style="background:#f0f0f0;"|'''''' | |||
| align="center" style="background:#f0f0f0;"|'''''' | |||
| align="center" style="background:#f0f0f0;"|'''''' | |||
| align="center" style="background:#f0f0f0;"|'''''' | |||
| align="center" style="background:#f0f0f0;"|'''''' | |||
| align="center" style="background:#f0f0f0;"|'''''' | |||
|- | |||
| high salt||||stock conc||||desired conc||vol|||||| | |||
|- | |||
| ||NaCl||5||M||2.64||50||mL||26.4||mL | |||
|- | |||
| ||KCl||4||M||0.11||50||mL||1.375||mL | |||
|- | |||
| ||water||||||||||||22.225||mL | |||
|- | |||
| low salt|||||||||||||||| | |||
|- | |||
| ||NaCl||5||M||0.264||50||mL||2.64||mL | |||
|- | |||
| ||KCl||4||M||0.011||50||mL||0.1375||mL | |||
|- | |||
| ||water||||||||||||47.2225||mL | |||
|- | |||
| | |||
|} | |||
==References== | ==References== | ||
<biblio> | <biblio> |
Revision as of 13:07, 13 October 2009
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In the beginning there was Jackgetting started here... The background reading I have done that will weigh heavy today: The Pierce Refolding Kit Instructions[1] that detail the 9 buffer components in their kit, which are: reducing agents A paper on High throughput automated refolding [2] that describes a 96 well matrix of buffer conditions they developed for structural genomics projects for Mtb and Viruses.
Designing Our Matrix
additionally, So, our matrix will include an expansion of pH, and various detergents around that may or may not help (it's a screen for a reason)
additives that are not NDSB's that may help, and some other stuff I like to hypothesize about. Stock Solutionsit is important to make sure that your stocks are at concentrations appropriate such that an all positive sample (e.g. one having all possible components) does not have a volume greater than your total volume for the experiment
NB: I used the totally wicked excel2wiki converter for the table code
Buffers=
salt mix
References
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