Editing LCC 500-501 Operation (section)

== Methods<br>  ==

Old FPLC in cold Room<br> 

Method 4: Load Protein to SP FF column (valve 3.3 &amp; 4.3), wash, then elute directly to Ni resin, wash and elute His tag protein<br> 

{| width="533" border="1" cellspacing="1" cellpadding="1"
|-
| Volume/ml<br> 
| Function<br> 
| Value<br> 
| Note<br>
|-
| 0.00<br> 
| VALVE.POS<br> 
| 1.2<br> 
| Inject<br>
|-
| 0.00<br> 
| VALVE.POS<br> 
| 2.1<br> 
| H2O<br>
|-
| 0.00<br> 
| VALVE.POS<br> 
| 5.1<br> 
| valve 5 by pass<br>
|-
| 0.00<br> 
| VALVE.POS<br> 
| 6.1<br> 
| valve 6&nbsp; by pass<br>
|-
| 0.00<br> 
| VALVE.POS<br> 
| 3.3<br> 
| SP FF<br>
|-
| 0.00<br> 
| VALVE.POS<br> 
| 4.3<br> 
| SP FF<br>
|-
| 0.00<br> 
| FLOW C<br> 
| 2.00<br> 
| Load Sample with P1<br>
|-
| 50<br> 
| ML/MIN<br> 
| 1.00<br> 
| sample volume <br>
|-
| 40.10<br> 
| ML/MIN<br> 
| 2.0<br> 
| start system wash (pump A, valves, tubing)<br>
|-
| 40.10<br> 
| CONC&nbsp;%B<br> 
| 0.00<br> 
| use water<br>
|-
| 75.10<br> 
| ML/MIN<br> 
| 0.00<br> 
| end wash<br>
|-
| 75.10<br> 
| WASH A.B<br> 
| 1.1<br> 
| <br>
|-
| 75.10 
| VALVE.POS 
| 1.1 
| <br>
|}

<br> Example Methods 

Method # 7: Load 10 ml sample using P1 pump<br> 

{| width="464" border="1" cellspacing="1" cellpadding="1"
|-
| Volume/ml<br> 
| Function<br> 
| Value<br> 
| Note<br>
|-
| 0.00<br> 
| VALVE.POS<br> 
| 2.2<br> 
| Buffer A<br>
|-
| 0.00<br> 
| VALVE.POS<br> 
| 4.1<br> 
| valve 4 by pass<br>
|-
| 0.00<br> 
| VALVE.POS<br> 
| 5.1<br> 
| valve 5 by pass<br>
|-
| 0.00<br> 
| VALVE.POS<br> 
| 6.1<br> 
| valve 6 waste<br>
|-
| 0.00<br> 
| VALVE.POS<br> 
| 3.1<br> 
| Flow to position 1 valve 3<br>
|-
| 0.00<br> 
| VALVE.POS<br> 
| 1.1<br> 
| switch MV-7 valve to Load<br>
|-
| 0.10<br> 
| FLOW C<br> 
| 2.00<br> 
| Load sample to superloop at 2 ml/min using P1 pump<br>
|-
| 10.10<br> 
| FLOW C<br> 
| 0.00<br> 
| stop pumping<br>
|-
| 10.10<br> 
| VALVE.POS<br> 
| 1.2<br> 
| switch valve to inject for sample injection<br>
|-
| 10.10<br> 
| ML/MIN<br> 
| 1.0<br> 
| inject sample<br>
|-
| 10.10<br> 
| CONC&nbsp;%B<br> 
| 0.00<br> 
| use Buffer A<br>
|-
| 20.10<br> 
| ML/MIN<br> 
| 0.00<br> 
| end sample injection<br>
|}

<br> 

Method # 0: System Wash<br> 

<br> 

{| width="533" border="1" cellspacing="1" cellpadding="1"
|-
| Volume/ml<br> 
| Function<br> 
| Value<br> 
| Note<br>
|-
| 0.00<br> 
| VALVE.POS<br> 
| 2.1<br> 
| Wash with water<br>
|-
| 0.00<br> 
| VALVE.POS<br> 
| 4.1<br> 
| valve 4 by pass<br>
|-
| 0.00<br> 
| VALVE.POS<br> 
| 5.1<br> 
| valve 5 by pass<br>
|-
| 0.00<br> 
| VALVE.POS<br> 
| 6.1<br> 
| valve 6 waste<br>
|-
| 0.00<br> 
| VALVE.POS<br> 
| 3.1<br> 
| Flow to position 1 valve 3<br>
|-
| 0.00<br> 
| VALVE.POS<br> 
| 1.3<br> 
| switch MV-7 valve to wash<br>
|-
| 0.10<br> 
| FLOW C<br> 
| 2.00<br> 
| Wash superloop at 2 ml/min using P1 pump<br>
|-
| 40.10<br> 
| FLOW C<br> 
| 0.00<br> 
| stop pumping<br>
|-
| 40.10<br> 
| ML/MIN<br> 
| 2.0<br> 
| start system wash (pump A, valves, tubing)<br>
|-
| 40.10<br> 
| CONC&nbsp;%B<br> 
| 0.00<br> 
| use water<br>
|-
| 75.10<br> 
| ML/MIN<br> 
| 0.00<br> 
| end wash<br>
|-
| 75.10<br> 
| WASH A.B<br> 
| 1.1<br> 
| <br>
|-
| 75.10 
| VALVE.POS 
| 1.1 
| <br>
|}

<br> 

Method # 1: Editable method (example)<br> 

{| width="533" border="1" cellspacing="1" cellpadding="1"
|-
| Volume/ml<br> 
| Function<br> 
| Value<br> 
| Note<br>
|-
| 0.00<br> 
| ML/MIN<br> 
| 0.5<br> 
| <br>
|-
| 0.00<br> 
| LOOP TMS<br> 
| 8<br> 
| Change this number according the sample volume, if it is 80 ml, loop 8 times, if it is 100 ml, loop 10 times<br>
|-
| 0.00<br> 
| CALL METH<br> 
| 7<br> 
| Load Sample using 10 ml superloop and P-1 pump<br>
|-
| 0.00<br> 
| END OF LOOP<br> 
| <br> 
| <br>
|-
| 0.00<br> 
| CALL METH<br> 
| 8<br> 
| &nbsp; Purify protein on 5 ml HisTrap HP column <br>
|}

<br> Method # 8: Purify His Tagged Protein using 5 ml HisTrap HP cartridge.<br> 

{| width="494" border="1" cellspacing="1" cellpadding="1"
|-
| Volume/ml<br> 
| Function<br> 
| Value<br> 
| Note<br>
|-
| 0.00<br> 
| CONC&nbsp;%B<br> 
| 0.0<br> 
| <br>
|-
| 0.00<br> 
| ML/MIN<br> 
| 1.00<br> 
| 1 ml/min<br>
|-
| 0.00<br> 
| CLEAR DATA<br> 
| <br> 
| <br>
|-
| 50.00<br> 
| CONC&nbsp;%B<br> 
| 4.0<br> 
| 20 mM&nbsp; (0.04X500 mM=20 mM)<br>
|-
| 100.00<br> 
| CONC&nbsp;%B<br> 
| 8.0<br> 
| 40 mM <br>
|-
| 150.00<br> 
| PORT.SET<br> 
| 6.1<br> 
| start fraction collector<br>
|-
| 150.00<br> 
| VALVE.POS<br> 
| 6.2<br> 
| set valve 6 to position 2<br>
|-
| 150.00<br> 
| CONC&nbsp;%B<br> 
| 10.0<br> 
| 50 mM<br>
|-
| 200.00<br> 
| CONC&nbsp;%B<br> 
| 60.0<br> 
| 300 mM<br>
|-
| 250.00<br> 
| CONC&nbsp;%B<br> 
| 0<br> 
| &nbsp;back to 10 mM<br>
|-
| 250.00<br> 
| PORT.SET<br> 
| 6.0<br> 
| stop collecting fractions<br>
|-
| 265.00<br> 
| ML/MIN<br> 
| 0<br> 
| <br>
|}

[[Category:Protocols]]