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(12 intermediate revisions by the same user not shown) |
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| | colspan="2"| | | | colspan="2"| |
| <!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | | <!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> |
| ==In the beginning there was Jack== | | =Refolding Matrix design and use= |
| getting started here... The background reading I have done that will weigh heavy today: | | getting started here... The background reading I have done that will weigh heavy today: |
| The Pierce Refolding Kit Instructions<cite>Pierce</cite> that detail the 9 buffer components in their kit, which are: | | The Pierce Refolding Kit Instructions<cite>Pierce</cite> that detail the 9 buffer components in their kit, which are: |
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| ==Designing Our Matrix== | | ==Designing Our Matrix== |
| <BR> | | <BR> |
| The problem with any matrix report, is that they are packed full of misc. chemicals that are either not in house, or prohibitively expensive... it is my experience to find a way to furnish results that are desired first, then ask for $$$ chemicals to improve future experiments second... | | The problem with any report on buffer matricies is that they are packed full of miscellaneous chemicals that are either not in house, or prohibitively expensive... it is my experience (professor management theory) that the best way to get expensive materials/equpment to support a new method/tehcnique is to furnish results that are desired but were only accessible from that new method/technique (grunt work), and then ask for $$$ chemicals to improve future experiments...<BR> |
| <BR> | |
| This excludes:<BR><BR> | | This excludes:<BR><BR> |
| <BR>all Non Detergent SulfoBetaines (NDSB's) that can or cannot be important | | <BR>all Non Detergent SulfoBetaines (NDSB's) that can or cannot be important |
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| 7.5 HEPES<BR> | | 7.5 HEPES<BR> |
| 8.2 TRIS<BR> | | 8.2 TRIS<BR> |
| 9.0 CHES<BR> | | 9.2 CHES<BR> |
| <BR> | | <BR> |
| I replaced BORATE pH 9.5 from <cite>Willis</cite> with CHES to alleviate complications that will arise during reagent preparation, since borate is only marginally soluble in water, and cannot be prepared as a 10X stock (500 mM).<BR><BR> | | I replaced BORATE pH 9.5 from <cite>Willis</cite> with CHES to alleviate complications that will arise during reagent preparation, since borate is only marginally soluble in water, and cannot be prepared as a 10X stock (500 mM).<BR><BR> |
| <BR> | | <BR> |
| <u>detergents/ concentration</u><BR> | | <u>detergents</u><BR> |
| triton X 100<BR> | | triton X 100<BR> |
| tween 80<BR> | | tween 80<BR> |
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| <BR>''NB: NONE is a category for all subsequent categories, because it must be included for matrix generation'' | | <BR>''NB: NONE is a category for all subsequent categories, because it must be included for matrix generation'' |
| <BR><BR> | | <BR><BR> |
| <u>reducing agents/ concentration</u><BR> | | <u>reducing agents</u><BR> |
| DTT<BR> | | DTT<BR> |
| TCEP<BR> | | TCEP<BR> |
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| <BR> | | <BR> |
| <BR> | | <BR> |
| Why BSA? this is my screen, and a I want to try something wacky.<BR>
| | Every additive looks reasonable except for 1... BSA. <BR>And, why BSA? this is my screen, and a I want to try something wacky.<BR><BR> |
| ===Stock Solutions===
| | ==Stock Solutions== |
| it 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 | | it 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 the total volume for the experiment...and that all components are soluble at the concentration determined here for the stock solution. |
| <BR><BR> | | <BR><BR> |
| here is the table I cooked up to figure out what "X" concentration I would need to make everything work, taking into account certain characteristics (e.g. if 20% glycerol will be the final concentration, then i cannot achieve more than a 5X stock solution) | | here is the table I cooked up to figure out what "X" concentration I would need to make everything work, taking into account certain characteristics (e.g. if 20% glycerol will be the final concentration, then i cannot achieve more than a 5X stock solution) |
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| |- | | |- |
| | Reducing agent||100X||1 | | | Reducing agent||100X||1 |
| |-
| |
| | ||||
| |
| |- | | |- |
| | ||total||93 | | | ||total||93 |
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| | Reducing agent||5||mM|| | | | Reducing agent||5||mM|| |
| |- | | |- |
| | | | | Divalent cation||2||mM|| |
| | |- |
| | | |
| |} | | |} |
|
| |
|
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| |- | | |- |
| | Reducing agent||500||mM | | | Reducing agent||500||mM |
| | |- |
| | | Divalent cation||200||mM |
| |- | | |- |
| | | | | |
| |} | | |} |
| ==Buffers== | | |
| | ===Buffers=== |
| {| {{table}} | | {| {{table}} |
| | align="center" style="background:#f0f0f0;"|'''Buffer''' | | | align="center" style="background:#f0f0f0;"|'''Buffer''' |
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| | | | | |
| |} | | |} |
| ==salt mix== | | ===salt mix=== |
| {| {{table}} | | {| {{table}} |
| | align="center" style="background:#f0f0f0;"|'''high salt''' | | | align="center" style="background:#f0f0f0;"|'''high salt''' |
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| |} | | |} |
|
| |
|
| ==divalent cation mix== | | ===divalent cation mix=== |
| {| {{table}} | | {| {{table}} |
| | align="center" style="background:#f0f0f0;"|'''divalent cations''' | | | align="center" style="background:#f0f0f0;"|'''divalent cations''' |
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| |} | | |} |
|
| |
|
| ==detergents== | | ===detergents=== |
|
| |
|
| {| {{table}} | | {| {{table}} |
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| =Matrix= | | =Matrix= |
| the first matrix can be found [http://openwetware.org/wiki/User:Timothy_L._Foley/Notebook/refolding_matrix/2009/10/13/Refolding_Matrix-10-13-2009 here] | | the first matrix can be found [http://openwetware.org/wiki/User:Timothy_L._Foley/Notebook/refolding_matrix/2009/10/13/Refolding_Matrix-10-13-2009 here] |
| The above conditions were paired in "Custom Design" of JMP-IN as follows: | | The above conditions were paired in "Custom Design" of JMP-IN as follows:<br> |
| pH: 5 level Categorical | | pH: 5 level Categorical<br> |
| | Detergent: 4 categories (none, triton, tween, chaps)<br> |
| | amino acids: 3 categories (none, arg, glu)<br> |
| | salts: none low high<br> |
| | crowders: none PEG Glycerol <br> |
| | divalent cation: 0, 1 <br> |
| | GnCl: : 0, 1 <br> |
| | BSA: 0, 1 <br> |
| | Ligand: 0, 1 <br> |
| | Reducing agent: 0, 1 <br> |
|
| |
|
| {| {{table}}
| |
| | align="center" style="background:#f0f0f0;"|'''Buffer #'''
| |
| | align="center" style="background:#f0f0f0;"|'''pH'''
| |
| | align="center" style="background:#f0f0f0;"|'''detergent'''
| |
| | align="center" style="background:#f0f0f0;"|'''amino acids'''
| |
| | align="center" style="background:#f0f0f0;"|'''salt'''
| |
| | align="center" style="background:#f0f0f0;"|'''crowders'''
| |
| | align="center" style="background:#f0f0f0;"|'''divalent cation'''
| |
| | align="center" style="background:#f0f0f0;"|'''GnCl'''
| |
| | align="center" style="background:#f0f0f0;"|'''BSA'''
| |
| | align="center" style="background:#f0f0f0;"|'''Ligand'''
| |
| | align="center" style="background:#f0f0f0;"|'''reducing agent'''
| |
| | align="center" style="background:#f0f0f0;"|'''Y'''
| |
| |-
| |
| | 1||5.5 MES||none||none||low||PEG||1||0||1||0||DTT||
| |
| |-
| |
| | 2||5.5 MES||none||none||high||none||0||0||0||0||DTT||
| |
| |-
| |
| | 3||5.5 MES||none||none||high||PEG||0||0||0||1||none||
| |
| |-
| |
| | 4||5.5 MES||none||Arg||none||PEG||1||0||0||1||TCEP||
| |
| |-
| |
| | 5||5.5 MES||none||Glut||low||Glycerol||1||0||1||0||TCEP||
| |
| |-
| |
| | 6||5.5 MES||TritonX||Arg||none||Glycerol||1||0||1||1||none||
| |
| |-
| |
| | 7||5.5 MES||TritonX||Arg||high||none||0||0||1||0||TCEP||
| |
| |-
| |
| | 8||5.5 MES||TritonX||Glut||none||PEG||0||0||1||0||DTT||
| |
| |-
| |
| | 9||5.5 MES||TritonX||Glut||low||none||0||1||0||1||DTT||
| |
| |-
| |
| | 10||5.5 MES||TritonX||Glut||low||Glycerol||0||1||0||1||none||
| |
| |-
| |
| | 11||5.5 MES||Tween80||none||low||Glycerol||1||1||1||0||TCEP||
| |
| |-
| |
| | 12||5.5 MES||Tween80||Arg||low||none||0||0||0||0||DTT||
| |
| |-
| |
| | 13||5.5 MES||Tween80||Arg||high||Glycerol||0||1||1||1||TCEP||
| |
| |-
| |
| | 14||5.5 MES||Tween80||Glut||none||Glycerol||1||1||0||1||TCEP||
| |
| |-
| |
| | 15||5.5 MES||Tween80||Glut||high||PEG||0||1||1||1||none||
| |
| |-
| |
| | 16||5.5 MES||CHAPS||none||low||none||0||1||0||0||none||
| |
| |-
| |
| | 17||5.5 MES||CHAPS||none||high||none||0||1||1||1||TCEP||
| |
| |-
| |
| | 18||5.5 MES||CHAPS||none||high||none||1||1||0||1||DTT||
| |
| |-
| |
| | 19||5.5 MES||CHAPS||Arg||none||PEG||1||0||0||0||none||
| |
| |-
| |
| | 20||5.5 MES||CHAPS||Glut||none||Glycerol||1||1||1||0||DTT||
| |
| |-
| |
| | 21||6.5 MES||none||none||none||none||1||0||0||1||none||
| |
| |-
| |
| | 22||6.5 MES||none||none||none||Glycerol||0||1||1||0||DTT||
| |
| |-
| |
| | 23||6.5 MES||none||Arg||low||none||1||1||1||1||none||
| |
| |-
| |
| | 24||6.5 MES||none||Arg||high||PEG||1||1||1||0||none||
| |
| |-
| |
| | 25||6.5 MES||none||Glut||low||Glycerol||1||0||1||1||DTT||
| |
| |-
| |
| | 26||6.5 MES||TritonX||none||none||Glycerol||1||0||0||0||TCEP||
| |
| |-
| |
| | 27||6.5 MES||TritonX||Arg||low||PEG||0||1||1||1||DTT||
| |
| |-
| |
| | 28||6.5 MES||TritonX||Arg||high||Glycerol||0||0||0||0||DTT||
| |
| |-
| |
| | 29||6.5 MES||TritonX||Glut||low||none||1||1||0||0||TCEP||
| |
| |-
| |
| | 30||6.5 MES||TritonX||Glut||high||Glycerol||1||1||0||1||none||
| |
| |-
| |
| | 31||6.5 MES||Tween80||none||none||none||0||1||1||1||DTT||
| |
| |-
| |
| | 32||6.5 MES||Tween80||none||low||PEG||1||0||0||0||TCEP||
| |
| |-
| |
| | 33||6.5 MES||Tween80||none||low||PEG||1||0||0||1||DTT||
| |
| |-
| |
| | 34||6.5 MES||Tween80||Arg||none||PEG||0||1||1||0||TCEP||
| |
| |-
| |
| | 35||6.5 MES||Tween80||Glut||high||none||0||0||1||0||DTT||
| |
| |-
| |
| | 36||6.5 MES||CHAPS||none||high||Glycerol||0||1||1||0||TCEP||
| |
| |-
| |
| | 37||6.5 MES||CHAPS||Arg||high||PEG||1||1||1||0||none||
| |
| |-
| |
| | 38||6.5 MES||CHAPS||Arg||high||Glycerol||0||0||1||1||none||
| |
| |-
| |
| | 39||6.5 MES||CHAPS||Glut||none||none||1||1||0||1||none||
| |
| |-
| |
| | 40||6.5 MES||CHAPS||Glut||low||PEG||0||0||0||1||TCEP||
| |
| |-
| |
| | 41||7.5 HEPES||none||none||low||PEG||0||1||0||1||none||
| |
| |-
| |
| | 42||7.5 HEPES||none||Arg||high||none||0||0||1||1||TCEP||
| |
| |-
| |
| | 43||7.5 HEPES||none||Arg||high||Glycerol||0||1||0||0||none||
| |
| |-
| |
| | 44||7.5 HEPES||none||Glut||none||PEG||0||1||0||0||TCEP||
| |
| |-
| |
| | 45||7.5 HEPES||none||Glut||high||Glycerol||0||0||0||0||DTT||
| |
| |-
| |
| | 46||7.5 HEPES||TritonX||none||none||none||0||0||1||1||none||
| |
| |-
| |
| | 47||7.5 HEPES||TritonX||none||none||none||1||0||1||0||TCEP||
| |
| |-
| |
| | 48||7.5 HEPES||TritonX||Arg||low||PEG||0||0||0||1||TCEP||
| |
| |-
| |
| | 49||7.5 HEPES||TritonX||Arg||low||Glycerol||1||1||1||0||none||
| |
| |-
| |
| | 50||7.5 HEPES||TritonX||Glut||high||PEG||1||1||0||0||TCEP||
| |
| |-
| |
| | 51||7.5 HEPES||Tween80||none||none||Glycerol||1||0||1||1||none||
| |
| |-
| |
| | 52||7.5 HEPES||Tween80||none||low||PEG||0||1||1||1||DTT||
| |
| |-
| |
| | 53||7.5 HEPES||Tween80||none||low||PEG||1||1||1||1||TCEP||
| |
| |-
| |
| | 54||7.5 HEPES||Tween80||Glut||none||none||1||1||1||1||DTT||
| |
| |-
| |
| | 55||7.5 HEPES||Tween80||Glut||high||none||1||0||0||1||DTT||
| |
| |-
| |
| | 56||7.5 HEPES||CHAPS||none||low||Glycerol||0||1||0||0||DTT||
| |
| |-
| |
| | 57||7.5 HEPES||CHAPS||Arg||none||Glycerol||1||1||0||0||TCEP||
| |
| |-
| |
| | 58||7.5 HEPES||CHAPS||Arg||low||none||0||0||1||1||none||
| |
| |-
| |
| | 59||7.5 HEPES||CHAPS||Glut||high||PEG||1||0||1||0||DTT||
| |
| |-
| |
| | 60||8.2 Tris||none||none||high||PEG||0||0||1||1||TCEP||
| |
| |-
| |
| | 61||8.2 Tris||none||Arg||none||none||1||1||1||0||DTT||
| |
| |-
| |
| | 62||8.2 Tris||none||Glut||none||none||0||0||1||1||none||
| |
| |-
| |
| | 63||8.2 Tris||none||Glut||low||none||0||1||0||0||TCEP||
| |
| |-
| |
| | 64||8.2 Tris||none||Glut||low||Glycerol||1||1||1||0||DTT||
| |
| |-
| |
| | 65||8.2 Tris||TritonX||none||low||PEG||1||1||1||1||none||
| |
| |-
| |
| | 66||8.2 Tris||TritonX||none||high||Glycerol||1||0||1||0||none||
| |
| |-
| |
| | 67||8.2 Tris||TritonX||Arg||high||PEG||1||1||0||1||DTT||
| |
| |-
| |
| | 68||8.2 Tris||TritonX||Glut||high||PEG||0||1||1||1||TCEP||
| |
| |-
| |
| | 69||8.2 Tris||Tween80||none||high||none||1||1||0||0||none||
| |
| |-
| |
| | 70||8.2 Tris||Tween80||Arg||low||Glycerol||0||1||0||0||DTT||
| |
| |-
| |
| | 71||8.2 Tris||Tween80||Arg||high||Glycerol||1||0||0||1||TCEP||
| |
| |-
| |
| | 72||8.2 Tris||Tween80||Glut||none||Glycerol||0||0||0||1||none||
| |
| |-
| |
| | 73||8.2 Tris||CHAPS||none||low||none||1||0||0||0||none||
| |
| |-
| |
| | 74||8.2 Tris||CHAPS||none||low||Glycerol||0||0||1||1||TCEP||
| |
| |-
| |
| | 75||8.2 Tris||CHAPS||Arg||none||none||0||1||0||0||DTT||
| |
| |-
| |
| | 76||8.2 Tris||CHAPS||Arg||none||PEG||1||0||0||1||DTT||
| |
| |-
| |
| | 77||8.2 Tris||CHAPS||Glut||none||PEG||0||0||1||0||TCEP||
| |
| |-
| |
| | 78||9.2 CHES||none||none||none||Glycerol||1||1||0||1||TCEP||
| |
| |-
| |
| | 79||9.2 CHES||none||Arg||none||PEG||0||1||1||0||none||
| |
| |-
| |
| | 80||9.2 CHES||none||Arg||low||none||1||1||0||1||TCEP||
| |
| |-
| |
| | 81||9.2 CHES||none||Glut||high||Glycerol||1||0||0||1||DTT||
| |
| |-
| |
| | 82||9.2 CHES||TritonX||none||none||none||0||0||0||0||TCEP||
| |
| |-
| |
| | 83||9.2 CHES||TritonX||none||none||Glycerol||0||1||1||1||DTT||
| |
| |-
| |
| | 84||9.2 CHES||TritonX||none||high||PEG||1||0||1||0||DTT||
| |
| |-
| |
| | 85||9.2 CHES||TritonX||Arg||low||none||1||1||1||1||DTT||
| |
| |-
| |
| | 86||9.2 CHES||TritonX||Glut||low||none||0||0||0||0||none||
| |
| |-
| |
| | 87||9.2 CHES||Tween80||none||high||PEG||0||1||0||0||none||
| |
| |-
| |
| | 88||9.2 CHES||Tween80||Arg||low||none||1||0||1||0||TCEP||
| |
| |-
| |
| | 89||9.2 CHES||Tween80||Arg||low||Glycerol||0||0||0||0||none||
| |
| |-
| |
| | 90||9.2 CHES||Tween80||Glut||none||PEG||0||1||0||0||none||
| |
| |-
| |
| | 91||9.2 CHES||Tween80||Glut||high||none||1||0||1||0||none||
| |
| |-
| |
| | 92||9.2 CHES||CHAPS||none||high||Glycerol||1||1||0||1||DTT||
| |
| |-
| |
| | 93||9.2 CHES||CHAPS||Arg||none||PEG||0||0||0||1||DTT||
| |
| |-
| |
| | 94||9.2 CHES||CHAPS||Glut||low||PEG||1||0||1||0||none||
| |
| |-
| |
| | 95||9.2 CHES||CHAPS||Glut||low||Glycerol||0||0||1||1||TCEP||
| |
| |-
| |
| | 96||9.2 CHES||CHAPS||Glut||high||none||1||1||1||1||TCEP||
| |
| |-
| |
| |
| |
| |}
| |
|
| |
|
| | the second matrix matrix can be found [http://openwetware.org/wiki/User:Timothy_L._Foley/Notebook/refolding_matrix/2009/10/13/Refolding_Matrix-10-13-2009_amino_acids_4-factor here] |
| | or the following matrix with the amino acids as a 4-factor categorical with (none, glu, arg, glu+arg) |
| | <br> |
| | <br> |
| | I am partial to exploring the largest number of possible buffer configurations, but I want it to still be worth while, i.e. if one of these randomized ingredients has negative effects on folding, a large portion of the wells is worthless. The software wants to prepare 480 wells as a default for a single iteration of the experiment... this is too many... I set the value at 96.. |
|
| |
|
| or the following matrix with the amino acids as a 4-factor categorical with none, glu, arg, glu+arg
| | ----<BR><BR> |
| | | it took all afternoon to prep the stock solutions above. Neither pH meter probe works, and we do not have any solid TCEP.<BR> I will go to the Core tomorrow and acquire some fresh DTT and TCEP.<BR> |
| {| {{table}}
| | '''NOTE: both amino acid solutions were adjusted to be 1/2 the concentration here. 58g Arg in 50 mL water (116% w/v solution) seemed a bit over the top.''' |
| | align="center" style="background:#f0f0f0;"|'''buffer #'''
| |
| | align="center" style="background:#f0f0f0;"|'''pH'''
| |
| | align="center" style="background:#f0f0f0;"|'''detergent'''
| |
| | align="center" style="background:#f0f0f0;"|'''amino acids'''
| |
| | align="center" style="background:#f0f0f0;"|'''salt'''
| |
| | align="center" style="background:#f0f0f0;"|'''crowders'''
| |
| | align="center" style="background:#f0f0f0;"|'''divalent cation'''
| |
| | align="center" style="background:#f0f0f0;"|'''GnCl'''
| |
| | align="center" style="background:#f0f0f0;"|'''BSA'''
| |
| | align="center" style="background:#f0f0f0;"|'''Ligand'''
| |
| | align="center" style="background:#f0f0f0;"|'''reducing agent'''
| |
| | align="center" style="background:#f0f0f0;"|'''Amino acid'''
| |
| |-
| |
| | 1||5.5 MES||none||none||low||none||1||0||1||1||none||Arg
| |
| |-
| |
| | 2||5.5 MES||none||none||high||Glycerol||0||1||1||1||TCEP||Glu
| |
| |-
| |
| | 3||5.5 MES||none||Glut||none||none||0||0||0||0||none||Arg+Glu
| |
| |-
| |
| | 4||5.5 MES||none||Glut||low||Glycerol||0||0||0||0||TCEP||none
| |
| |-
| |
| | 5||5.5 MES||TritonX||none||high||none||1||0||0||1||none||Glu
| |
| |-
| |
| | 6||5.5 MES||TritonX||Arg||low||PEG||1||1||1||1||none||Arg+Glu
| |
| |-
| |
| | 7||5.5 MES||TritonX||Arg||low||Glycerol||0||1||0||0||TCEP||Arg
| |
| |-
| |
| | 8||5.5 MES||TritonX||Arg||high||Glycerol||1||1||1||0||none||Arg+Glu
| |
| |-
| |
| | 9||5.5 MES||TritonX||Glut||none||none||1||0||1||0||TCEP||Arg+Glu
| |
| |-
| |
| | 10||5.5 MES||Tween80||none||none||PEG||0||1||1||1||DTT||none
| |
| |-
| |
| | 11||5.5 MES||Tween80||none||none||Glycerol||1||1||0||0||DTT||Arg
| |
| |-
| |
| | 12||5.5 MES||Tween80||Arg||none||PEG||0||0||0||1||TCEP||Arg
| |
| |-
| |
| | 13||5.5 MES||Tween80||Arg||low||Glycerol||0||0||0||0||none||Glu
| |
| |-
| |
| | 14||5.5 MES||Tween80||Arg||high||none||1||1||1||1||DTT||Arg
| |
| |-
| |
| | 15||5.5 MES||CHAPS||none||high||Glycerol||0||0||0||0||DTT||Arg+Glu
| |
| |-
| |
| | 16||5.5 MES||CHAPS||Arg||none||none||1||0||0||1||TCEP||none
| |
| |-
| |
| | 17||5.5 MES||CHAPS||Glut||none||PEG||0||1||0||1||none||Glu
| |
| |-
| |
| | 18||5.5 MES||CHAPS||Glut||low||PEG||1||1||1||0||DTT||Glu
| |
| |-
| |
| | 19||5.5 MES||CHAPS||Glut||high||PEG||1||1||1||0||DTT||none
| |
| |-
| |
| | 20||6.5 MES||none||none||none||none||1||1||0||0||none||Arg
| |
| |-
| |
| | 21||6.5 MES||none||none||low||Glycerol||1||0||0||1||DTT||none
| |
| |-
| |
| | 22||6.5 MES||none||Arg||none||PEG||1||0||0||1||DTT||none
| |
| |-
| |
| | 23||6.5 MES||none||Glut||high||none||0||1||1||0||none||Arg+Glu
| |
| |-
| |
| | 24||6.5 MES||none||Glut||high||PEG||0||1||0||0||TCEP||Arg+Glu
| |
| |-
| |
| | 25||6.5 MES||TritonX||none||low||none||0||1||0||0||TCEP||none
| |
| |-
| |
| | 26||6.5 MES||TritonX||none||high||Glycerol||1||1||1||1||DTT||Arg
| |
| |-
| |
| | 27||6.5 MES||TritonX||Arg||none||none||0||1||0||0||none||Glu
| |
| |-
| |
| | 28||6.5 MES||TritonX||Arg||high||PEG||1||0||1||1||DTT||Arg+Glu
| |
| |-
| |
| | 29||6.5 MES||TritonX||Glut||low||Glycerol||1||0||1||1||none||Arg
| |
| |-
| |
| | 30||6.5 MES||Tween80||Arg||none||none||0||1||1||1||DTT||Arg
| |
| |-
| |
| | 31||6.5 MES||Tween80||Arg||none||Glycerol||0||0||1||1||none||Arg+Glu
| |
| |-
| |
| | 32||6.5 MES||Tween80||Glut||low||PEG||1||1||0||0||TCEP||Glu
| |
| |-
| |
| | 33||6.5 MES||Tween80||Glut||low||Glycerol||1||1||1||1||TCEP||none
| |
| |-
| |
| | 34||6.5 MES||Tween80||Glut||high||Glycerol||0||1||1||1||none||Glu
| |
| |-
| |
| | 35||6.5 MES||CHAPS||none||none||none||0||0||1||0||TCEP||Arg
| |
| |-
| |
| | 36||6.5 MES||CHAPS||none||low||PEG||0||0||1||0||none||Glu
| |
| |-
| |
| | 37||6.5 MES||CHAPS||none||low||Glycerol||1||0||0||0||TCEP||Arg+Glu
| |
| |-
| |
| | 38||6.5 MES||CHAPS||Arg||high||PEG||0||0||0||1||TCEP||none
| |
| |-
| |
| | 39||6.5 MES||CHAPS||Arg||high||PEG||1||0||0||0||DTT||Glu
| |
| |-
| |
| | 40||7.5 HEPES||none||none||high||none||1||0||0||1||TCEP||Glu
| |
| |-
| |
| | 41||7.5 HEPES||none||Arg||none||Glycerol||0||0||1||0||DTT||Glu
| |
| |-
| |
| | 42||7.5 HEPES||none||Glut||none||Glycerol||1||1||1||1||TCEP||Arg+Glu
| |
| |-
| |
| | 43||7.5 HEPES||none||Glut||low||none||0||0||1||1||DTT||none
| |
| |-
| |
| | 44||7.5 HEPES||none||Glut||low||PEG||0||0||0||1||DTT||Arg
| |
| |-
| |
| | 45||7.5 HEPES||TritonX||none||none||PEG||1||1||1||0||DTT||Glu
| |
| |-
| |
| | 46||7.5 HEPES||TritonX||none||low||none||0||1||0||1||DTT||Glu
| |
| |-
| |
| | 47||7.5 HEPES||TritonX||none||high||PEG||0||0||1||1||none||Arg
| |
| |-
| |
| | 48||7.5 HEPES||TritonX||Arg||none||PEG||0||0||1||0||none||none
| |
| |-
| |
| | 49||7.5 HEPES||TritonX||Glut||high||Glycerol||0||0||0||0||TCEP||Glu
| |
| |-
| |
| | 50||7.5 HEPES||Tween80||none||low||PEG||1||0||1||1||TCEP||Arg+Glu
| |
| |-
| |
| | 51||7.5 HEPES||Tween80||none||high||Glycerol||1||0||1||0||TCEP||Arg+Glu
| |
| |-
| |
| | 52||7.5 HEPES||Tween80||Arg||high||none||1||0||0||0||DTT||none
| |
| |-
| |
| | 53||7.5 HEPES||Tween80||Glut||none||PEG||1||1||0||0||TCEP||Arg+Glu
| |
| |-
| |
| | 54||7.5 HEPES||Tween80||Glut||high||none||1||1||0||0||none||Arg
| |
| |-
| |
| | 55||7.5 HEPES||CHAPS||none||none||none||0||1||1||1||none||none
| |
| |-
| |
| | 56||7.5 HEPES||CHAPS||Arg||none||Glycerol||1||1||0||0||none||none
| |
| |-
| |
| | 57||7.5 HEPES||CHAPS||Arg||low||none||0||1||1||0||DTT||Arg+Glu
| |
| |-
| |
| | 58||7.5 HEPES||CHAPS||Arg||low||Glycerol||1||1||0||1||TCEP||Arg
| |
| |-
| |
| | 59||7.5 HEPES||CHAPS||Glut||high||Glycerol||0||1||0||1||none||Arg
| |
| |-
| |
| | 60||8.2 Tris||none||none||none||PEG||0||1||0||0||TCEP||Arg
| |
| |-
| |
| | 61||8.2 Tris||none||none||none||Glycerol||0||1||1||0||none||none
| |
| |-
| |
| | 62||8.2 Tris||none||Arg||high||none||1||1||1||1||TCEP||Glu
| |
| |-
| |
| | 63||8.2 Tris||none||Arg||high||PEG||0||1||0||1||none||Arg+Glu
| |
| |-
| |
| | 64||8.2 Tris||none||Arg||high||PEG||1||0||0||0||DTT||Arg
| |
| |-
| |
| | 65||8.2 Tris||TritonX||none||low||PEG||0||0||0||1||none||Arg+Glu
| |
| |-
| |
| | 66||8.2 Tris||TritonX||Glut||none||PEG||1||1||1||0||TCEP||Arg
| |
| |-
| |
| | 67||8.2 Tris||TritonX||Glut||none||Glycerol||1||0||1||1||TCEP||none
| |
| |-
| |
| | 68||8.2 Tris||TritonX||Glut||low||none||1||1||0||0||DTT||none
| |
| |-
| |
| | 69||8.2 Tris||TritonX||Glut||high||Glycerol||0||0||0||1||DTT||none
| |
| |-
| |
| | 70||8.2 Tris||Tween80||none||low||none||0||0||1||0||DTT||Arg
| |
| |-
| |
| | 71||8.2 Tris||Tween80||none||high||none||0||0||0||0||none||none
| |
| |-
| |
| | 72||8.2 Tris||Tween80||Arg||low||PEG||0||1||1||1||TCEP||Glu
| |
| |-
| |
| | 73||8.2 Tris||Tween80||Arg||high||none||1||0||1||0||TCEP||Arg+Glu
| |
| |-
| |
| | 74||8.2 Tris||CHAPS||none||none||Glycerol||1||1||0||1||DTT||Arg+Glu
| |
| |-
| |
| | 75||8.2 Tris||CHAPS||Arg||low||Glycerol||0||1||1||0||DTT||Glu
| |
| |-
| |
| | 76||8.2 Tris||CHAPS||Glut||none||none||1||0||1||1||none||Glu
| |
| |-
| |
| | 77||8.2 Tris||CHAPS||Glut||low||Glycerol||1||0||0||1||none||Arg
| |
| |-
| |
| | 78||9.2 CHES||none||none||none||Glycerol||1||0||1||0||DTT||Glu
| |
| |-
| |
| | 79||9.2 CHES||none||Arg||low||none||0||1||0||1||TCEP||Arg+Glu
| |
| |-
| |
| | 80||9.2 CHES||none||Arg||low||PEG||1||0||1||0||none||Arg
| |
| |-
| |
| | 81||9.2 CHES||none||Arg||low||Glycerol||1||1||1||0||none||none
| |
| |-
| |
| | 82||9.2 CHES||none||Glut||high||none||1||1||1||1||DTT||Glu
| |
| |-
| |
| | 83||9.2 CHES||TritonX||none||high||Glycerol||0||1||1||0||TCEP||none
| |
| |-
| |
| | 84||9.2 CHES||TritonX||Arg||none||none||1||0||0||1||TCEP||Glu
| |
| |-
| |
| | 85||9.2 CHES||TritonX||Arg||high||Glycerol||0||1||0||0||DTT||Arg
| |
| |-
| |
| | 86||9.2 CHES||TritonX||Glut||none||PEG||0||1||0||1||DTT||Arg+Glu
| |
| |-
| |
| | 87||9.2 CHES||Tween80||none||low||PEG||1||1||0||1||none||none
| |
| |-
| |
| | 88||9.2 CHES||Tween80||none||high||PEG||1||1||0||0||none||Glu
| |
| |-
| |
| | 89||9.2 CHES||Tween80||Arg||none||Glycerol||1||0||0||1||none||Glu
| |
| |-
| |
| | 90||9.2 CHES||Tween80||Glut||none||Glycerol||0||0||0||1||DTT||Arg+Glu
| |
| |-
| |
| | 91||9.2 CHES||Tween80||Glut||low||none||0||0||1||0||DTT||Arg+Glu
| |
| |-
| |
| | 92||9.2 CHES||CHAPS||none||low||none||1||1||0||1||DTT||Arg+Glu
| |
| |-
| |
| | 93||9.2 CHES||CHAPS||none||high||PEG||0||0||1||1||TCEP||Arg
| |
| |-
| |
| | 94||9.2 CHES||CHAPS||Arg||high||none||0||1||1||1||TCEP||none
| |
| |-
| |
| | 95||9.2 CHES||CHAPS||Glut||none||none||0||0||1||0||TCEP||Arg
| |
| |-
| |
| | 96||9.2 CHES||CHAPS||Glut||high||PEG||1||0||1||0||none||none
| |
| |-
| |
| |
| |
| |}
| |
| | |
| ==References== | | ==References== |
| <biblio> | | <biblio> |