User contributions for Barkow

27 July 2009

• 21:4021:40, 27 July 2009 diff hist +12 Sauer:FtsH purification No edit summary current
• 21:3821:38, 27 July 2009 diff hist +8 Sauer:FtsH purification No edit summary
• 21:3721:37, 27 July 2009 diff hist +3,288 N Sauer:FtsH purification New page: =Wild-type FtsH Expression and Purification:= (From Herman et al. 2003) E.Coli strain JM105 pBHB1 Purification for 4L. MW ~ 71kD # Grow in TB/Mg2+ with Chloroamphenicol to OD600 0.6 at...
• 21:3421:34, 27 July 2009 diff hist +29 Sauer:Protocols →Protein Expression/Purification current
• 21:3321:33, 27 July 2009 diff hist −1 Sauer:GST-sFtsH purification No edit summary current
• 21:3321:33, 27 July 2009 diff hist −3 Sauer:GST-sFtsH purification No edit summary
• 21:3121:31, 27 July 2009 diff hist +39 Sauer:GST-sFtsH purification →GST-sFtsH purification
• 21:3021:30, 27 July 2009 diff hist +1,504 N Sauer:GST-sFtsH purification New page: {{Sauer lab sidebar}} ===GST-sFtsH purification=== 1) Inoculate overnight from glycerol stock. 2) Wash overnight growth (spin down, resuspend, repeat) before inoculating 2x1L TB/Amp 3) ...
• 21:2921:29, 27 July 2009 diff hist +34 Sauer:Protocols →Protein Expression/Purification

7 July 2009

• 19:0419:04, 7 July 2009 diff hist +228 Sauer:In vitro peptide degradation by ClpXP →Degradation of peptides containing Abz and Y(NO2) by ClpXP current
• 18:3018:30, 7 July 2009 diff hist +19 Sauer:In vitro peptide degradation by ClpXP No edit summary
• 18:2518:25, 7 July 2009 diff hist +2 Sauer:In vitro peptide degradation by ClpXP →Correlating Rates with Concentration
• 18:2418:24, 7 July 2009 diff hist +740 Sauer:In vitro peptide degradation by ClpXP No edit summary
• 18:0518:05, 7 July 2009 diff hist +1,990 N Sauer:In vitro peptide degradation by ClpXP New page: {{Sauer lab sidebar}} ==Protocol 1== For peptides containing Abz and Y(NO2) ===PD buffer (1X)=== 25 mM HEPES-KOH, pH 7.6 5 mM MgCl₂ 0.032% NP-40 (Nonidet P40 substitute) * ...
• 17:5917:59, 7 July 2009 diff hist +49 Sauer:Protocols →In vitro enzymatic assays

30 June 2009

• 17:3617:36, 30 June 2009 diff hist +12 Sauer:ClpA purification →Buffers current
• 17:3217:32, 30 June 2009 diff hist +19 Sauer:ClpA purification →Buffers
• 17:3117:31, 30 June 2009 diff hist +20 Sauer:ClpA purification No edit summary
• 17:2917:29, 30 June 2009 diff hist +2,892 N Sauer:ClpA purification New page: {{Sauer lab sidebar}} ClpA Purifcation Protocol Jon Kenniston / Julia Flynn / John Lo General Notes • Do all purification steps at 4oC. • Thoroughly wash columns before us...
• 17:2917:29, 30 June 2009 diff hist +29 Sauer:Protocols No edit summary

23 June 2009

• 16:2216:22, 23 June 2009 diff hist +5 Sauer:FPLC protocol No edit summary current
• 16:2116:21, 23 June 2009 diff hist +21 Sauer:FPLC protocol No edit summary
• 16:2016:20, 23 June 2009 diff hist +1,836 N Sauer:FPLC protocol New page: == How to use the FPLC == '''Preparation:''' # Sign up for FPLC. # Prepare H2O and 20% EtOH (unless there is already plenty) ## Both must be filtered and degassed. # Prepare buffers ##...
• 16:1816:18, 23 June 2009 diff hist +1 Sauer:Protocols →Other good stuff
• 16:1816:18, 23 June 2009 diff hist +24 Sauer:Protocols →Other good stuff
• 16:1816:18, 23 June 2009 diff hist +22 Sauer:HPLC protocol No edit summary current
• 16:1616:16, 23 June 2009 diff hist 0 Sauer:HPLC protocol No edit summary
• 16:1616:16, 23 June 2009 diff hist +4 Sauer:HPLC protocol No edit summary
• 16:1516:15, 23 June 2009 diff hist −2 Sauer:HPLC protocol No edit summary
• 16:1416:14, 23 June 2009 diff hist −22 Sauer:HPLC protocol No edit summary
• 16:1316:13, 23 June 2009 diff hist 0 Sauer:HPLC protocol No edit summary
• 16:1216:12, 23 June 2009 diff hist +1 Sauer:HPLC protocol No edit summary
• 16:1216:12, 23 June 2009 diff hist −3 Sauer:HPLC protocol No edit summary
• 16:1116:11, 23 June 2009 diff hist +1 Sauer:HPLC protocol No edit summary
• 16:1116:11, 23 June 2009 diff hist +3 Sauer:HPLC protocol No edit summary
• 16:1016:10, 23 June 2009 diff hist +34 Sauer:HPLC protocol No edit summary
• 16:0916:09, 23 June 2009 diff hist +25 Sauer:HPLC protocol No edit summary
• 16:0816:08, 23 June 2009 diff hist +1,526 N Sauer:HPLC protocol New page: HPLC usage: Buffers: A: 0.06% TFA in dH2O. B: 0.054% TFA in 80% Acetonitrile When preparing, filter with Chemically Resistant ZapCap. 1) Turn on pumps from bottom to top. (there are 4 pa...
• 16:0716:07, 23 June 2009 diff hist +25 Sauer:Protocols No edit summary

18 June 2008

• 15:3215:32, 18 June 2008 diff hist +9 Sauer:GFP-H6-ssrA purification No edit summary current
• 15:3215:32, 18 June 2008 diff hist +74 Sauer:GFP-H6-ssrA purification No edit summary
• 15:3015:30, 18 June 2008 diff hist +4 Sauer:GFP-H6-ssrA purification No edit summary
• 15:2915:29, 18 June 2008 diff hist +16 Sauer:GFP-H6-ssrA purification No edit summary
• 15:2815:28, 18 June 2008 diff hist +17 Sauer:GFP-H6-ssrA purification No edit summary
• 15:2615:26, 18 June 2008 diff hist +25 Sauer:GFP-H6-ssrA purification No edit summary
• 15:2615:26, 18 June 2008 diff hist +6 Sauer:GFP-H6-ssrA purification No edit summary
• 15:2515:25, 18 June 2008 diff hist +233 Sauer:GFP-H6-ssrA purification No edit summary
• 15:2215:22, 18 June 2008 diff hist +435 N Sauer:GFP-H6-ssrA purification New page: Expression Grow cells at 30 C. Innoculate 1L of LB or TB + Amp with 10mL of overnight culture. Induce cells with 0.5mM IPTG at OD ~0.7 After ~ 3 hrs harvest cells by centrifugation, 20m ...
• 15:1315:13, 18 June 2008 diff hist +36 Sauer:Protocols →Protein Expression/Purification

7 November 2007

• 15:4615:46, 7 November 2007 diff hist 0 Sauer:His6 ClpX purification No edit summary current