User contributions for ESO wikiadmin

16 June 2025

• 13:1413:14, 16 June 2025 diff hist +10 AG Hauck - LabWiki No edit summary current Tag: Visual edit
• 13:1313:13, 16 June 2025 diff hist +15 Template:Labor-Boxes No edit summary current
• 13:1213:12, 16 June 2025 diff hist +1 Plasmid-Miniprep (Birnboim – Dooley protocol) No edit summary current Tag: Visual edit
• 13:1113:11, 16 June 2025 diff hist 0 Western Blot: semi-dry blot No edit summary current Tag: Visual edit
• 10:1910:19, 16 June 2025 diff hist +1 Lab Etiquette (ENG) →Lab Book current Tag: Visual edit
• 10:1910:19, 16 June 2025 diff hist −1 Lab Etiquette (ENG) No edit summary Tag: Visual edit

5 June 2025

• 14:4314:43, 5 June 2025 diff hist +74 Protein expression and purification No edit summary current Tag: Visual edit
• 14:4214:42, 5 June 2025 diff hist +33 Protein expression and purification →Concentration Determination of purified recombinant hPOPX2 (PPM1F) Tag: Visual edit
• 14:4214:42, 5 June 2025 diff hist +9 N File:SBA curve.png No edit summary current
• 14:4114:41, 5 June 2025 diff hist +45 Protein expression and purification →Quantification Gel for recombinant hPOPX2 (PPM1F) after Ulp1 treatment and dialysis – Gel #26 Tag: Visual edit
• 14:4014:40, 5 June 2025 diff hist +6 N File:SBA 25.png No edit summary current
• 14:3914:39, 5 June 2025 diff hist 0 Protein expression and purification →Purification post Ulp1 treatment for His-SUMO tag cleavage of His-SUMO hPOPX2 – Gel #24 – Part 2/2 Tag: Visual edit
• 14:3914:39, 5 June 2025 diff hist −8 Protein expression and purification →Quantification Gel for recombinant hPOPX2 (PPM1F) after Ulp1 treatment and dialysis – Gel #26 Tag: Visual edit
• 14:3814:38, 5 June 2025 diff hist +6 N File:SBA 24.png No edit summary current
• 14:3714:37, 5 June 2025 diff hist −36 Protein expression and purification No edit summary Tag: Visual edit
• 14:3714:37, 5 June 2025 diff hist −148 Protein expression and purification No edit summary Tag: Visual edit
• 14:3514:35, 5 June 2025 diff hist −48 Protein expression and purification No edit summary Tag: Visual edit
• 14:3414:34, 5 June 2025 diff hist +6 N File:SBA 21.png No edit summary current
• 14:3214:32, 5 June 2025 diff hist +9 Protein expression and purification No edit summary Tag: Visual edit
• 14:3114:31, 5 June 2025 diff hist +34 Protein expression and purification No edit summary Tag: Visual edit
• 14:3114:31, 5 June 2025 diff hist +5 N File:SB20.png No edit summary current
• 14:3014:30, 5 June 2025 diff hist −259 Protein expression and purification No edit summary Tag: Visual edit
• 14:2714:27, 5 June 2025 diff hist +7,452 N Protein expression and purification Created page with "== '''Expression of  HisSUMO-hPOPX2 (SAK) – Part 1/2''' == -      '''''Bacteria Sample'':''' #570 BL21 DE3 pRos. pET24a His-SUMO_hPopx2_fulllength -      One day prior, aprox. 15:00 : inoculate 50 mL of antibiotics containing media with 3 colonies and incubate o/n at 37°C -      7 mL each of overnight culture were used to inoculate 3 x 333 mL of antibiotics (Kan, cam) containing LB-medium in a 1 L Erlenmeyer flask until OD = 0.7 -      When OD is reach..." Tag: Visual edit
• 14:2214:22, 5 June 2025 diff hist +4 Lab Etiquette No edit summary current Tag: Visual edit
• 14:2014:20, 5 June 2025 diff hist +450 Testexpression in Pichia pastoris No edit summary current Tag: Visual edit
• 14:1714:17, 5 June 2025 diff hist +1,056 N Testexpression in Pichia pastoris Created page with " == Procedure == * Inoculate 5 mL YPD medium with one '''big''' colony ** in 50 ml falcon tube (secure lid with tape, don’t tighten lid) * Grow o. n. 30 °C, 200 rpm * Centrifuge at 2000 g, 5 min * Resuspend the complete pellet in 5 mL BMMY medium * Measure the OD ** Use a 1:100 dilution for measurement * Start growth ** For starters you don’t need to dilute your suspension ** For optimization purposes you may want to adjust your starting OD *** Recommended is a sta..." Tag: Visual edit
• 14:1014:10, 5 June 2025 diff hist +16 Transformation of Pichia pastoris No edit summary current Tag: Visual edit
• 14:0914:09, 5 June 2025 diff hist +32 Transformation of Pichia pastoris No edit summary Tag: Visual edit
• 14:0914:09, 5 June 2025 diff hist +428 Transformation of Pichia pastoris No edit summary Tag: Visual edit
• 14:0714:07, 5 June 2025 diff hist +1,023 N Transformation of Pichia pastoris Created page with " == Procedure == * Linearize 20 µg of plasmid of interest with EcoRV * Purify via EtOH precipitation: ** Adjust sample volume to 50 µL (MilliQ) ** Add 5 µL sodium acetate, vortex ** Add 125 µL ice cold EtOH abs.; Incubate on ice for 15 min ** Centrifuge 30 min, 14’000 rpm, 4 °C ** Discard the supernatant ** Wash the pellet with 500 µL colt EtOH 70 % ** Centrifuge 15 min, 14’000 rpm, 4 °C ** Remove the supernatant, dry pellet at RT ** Dissolve in 15-25 µL Mil..." Tag: Visual edit
• 14:0514:05, 5 June 2025 diff hist −7 Generation of competent Pichia pastoris No edit summary current Tag: Visual edit
• 14:0514:05, 5 June 2025 diff hist −130 Generation of competent Pichia pastoris No edit summary Tag: Visual edit
• 14:0414:04, 5 June 2025 diff hist +986 N Generation of competent Pichia pastoris Created page with "{{Protokoll-Mix|Inhalt=Wildtype Pichia Pastoris yJC100, grown on YPD agar plate *YPD medium (yeast extract (10 g/L), peptone (20 g/L), glucose (20 g/L)) *Transformation buffer (Tris (10 mM) pH 7,5, Sorbitol (0,6 M), LiAc (100mM), DTT (10 mM)) *Sorbitol (1M) *Inoculation loop *Erlenmeyer flask 500 mL *Spectrometer for OD measurement *50 mL falcons}} ===== Procedure ===== -       Inoculate 5 mL of YPD medium with one big colony -       Grow 8 h at 30 °C, 200 rpm..." Tag: Visual edit
• 14:0314:03, 5 June 2025 diff hist −3,053 Purification of GST-fusion proteins No edit summary current Tag: Visual edit
• 14:0314:03, 5 June 2025 diff hist +429 Purification of GST-fusion proteins No edit summary Tag: Visual edit
• 13:5913:59, 5 June 2025 diff hist +3,053 Purification of GST-fusion proteins No edit summary
• 13:5913:59, 5 June 2025 diff hist +687 Purification of GST-fusion proteins No edit summary Tag: Visual edit
• 13:5613:56, 5 June 2025 diff hist +35 Purification of GST-fusion proteins No edit summary Tag: Visual edit
• 13:5613:56, 5 June 2025 diff hist +1,178 N Purification of GST-fusion proteins Created page with "== Procedure == The frozen bacteria pellets are thawed in 5 ml T-buffer on ice and resuspended. Afterwards the samples are incubated with 1 ml lysozyme for 30 min. The cells are broken through the addition of 20 ml lysis buffer and the treatment of ultrasonic sound (impulse 50%, 20sec, 3-5x) on ice water. 300 µl sepharose-solution is added to the suspension and incubated for 10 min with head-over-head rotation. Afterwards the lysate is centrifuged for 20 min at 16000 r..." Tag: Visual edit
• 13:5313:53, 5 June 2025 diff hist +3 Expression of GST-fusion proteins No edit summary current Tag: Visual edit
• 13:5313:53, 5 June 2025 diff hist +2,000 N Expression of GST-fusion proteins Created page with "{{Protokoll-Mix|Inhalt=* *LB-Medium (10g Bacto-Trypton, 5g yeast extract, 5g NaCl, pH 7.0, ad 1l A. bidest) *Ampicillin (100 µg/ml) *IPTG (100 mM Isopropylthiogalactosid) *2xSDS sample buffer 125mM Tris-HCl (pH 6.8), 10% (v/v)-β- *Mercaptoethanol, 5% (w/v) SDS, 0.1% (w/v) Bromphenolblau, 20% (v/v) glycerin *T-buffer 100mM Tris (pH 8.0), 5 mM EDTA *GST-antibody}} == Procedure == === '''Test expression:''' === After cloning, the constructs of the desired GST-fusion pr..." Tag: Visual edit
• 13:5013:50, 5 June 2025 diff hist +1,616 N Bacterial Pulldown with soluble receptors Created page with "{{Protokoll-Mix|titel=Materials|Inhalt=PBS (1x) – 24g NaCl, 0.6g KCl, 3.45g Na2HPO4*7H2O, 0.6g KH2PO4, pH 7.4, ad 1l A.bidest<br>Cell culture supernatants (e.g. OptiMEM) harvested from 293cells transiently transfected with constructs encoding soluble CEACAM- ectodomains fused to GFP<br>Polyclonal rabbit anti-GFP antibody (AG Hauck)<br>2xSDS sample buffer 125mM Tris-HCl (pH 6.8), 10% (v/v) β-Mercaptoethanol, 5% (w/v) SDS, 0.1% (w/v) Bromphenolblau, 20% (v/v) Glycerin}}..." current Tag: Visual edit
• 13:4813:48, 5 June 2025 diff hist +599 N Opsonizing bacteria with Fc fusion proteins Created page with "Bacteria are stained with CFSE and resuspended in PBS. 0.3 bacteria (OD 550) are incubated with either 0.05 mg of purified fusion proteins of 1ml of the unpurified fusion protein for 1 h under rotation at 4°C. As a positive control bacteria are incubated with 500 µl of anti-gonococcal sera (heat-inactivated for 1 h at 56°C) and 500 µl PBS. As a negative control bacteria are incubated with 1 ml of PBS. After the incubation the bacteria are washed and resuspended in 1..." current Tag: Visual edit
• 13:2013:20, 5 June 2025 diff hist +125 Lab Etiquette (ENG) No edit summary Tag: Visual edit
• 13:1913:19, 5 June 2025 diff hist +43 Lab Etiquette (ENG) No edit summary Tag: Visual edit
• 13:1713:17, 5 June 2025 diff hist −6,081 Lab Etiquette No edit summary Tag: Visual edit
• 13:1413:14, 5 June 2025 diff hist +17 Lab Etiquette →Laborbuch Tag: Visual edit
• 12:5912:59, 5 June 2025 diff hist +184 Laborsicherheit No edit summary current Tag: Visual edit
• 12:5812:58, 5 June 2025 diff hist −2 All About Safety No edit summary current Tag: Visual edit
• 12:5812:58, 5 June 2025 diff hist +35 All About Safety No edit summary Tag: Visual edit