Summary of Contents for Bico Cellenion proteoCHIP EVO 96
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User Manual proteoCHIP EVO 96 for SCP User Manual proteoCHIP EVO 96 for SCP (Single Cell Proteomics) Rev.0.0 – eff. date: 07/12/2023 UM_proteoCHIP_05 Page: 1/14...
User Manual proteoCHIP EVO 96 for SCP Table of contents Document history ......................2 Introduction ......................3 Equipment and reagents ..................4 Equipment needed ................... 4 Reagents needed ..................... 5 Cell culture ....................... 5 Preparation of the cellenONE .................. 6 Preparation of the system ................
User Manual proteoCHIP EVO 96 for SCP 1 Introduction Important notes will be highlighted in green boxes. All basic functions of the cellenONE associated with dispensing or cell sorting are presented in great details in the User Manual for cellenONE. Furthermore, the User Manual for Proteomics Workflows provides step-by-step instructions with software screenshots through the most basic procedures referenced in this manual.
User Manual proteoCHIP EVO 96 for SCP 2.2 Reagents needed Use only MS-grade reagents and prepare dilutions with purified water (>18 MΩ, <3ppb TOC at 25 °C) See the full recipe in Chapter 4.1. • Master mix protease: 100 ng/µL Trypsin or Trypsin-LysC mixture in 50 mM acetic acid.
User Manual proteoCHIP EVO 96 for SCP 3 Preparation of the cellenONE 3.1 Preparation of the system • Exchange Water of the System Liquid bottle: Degas fresh MilliQ-Water for 15 min by applying a vacuum and placing the System Liquid bottle inside the ultrasonic water bath.
User Manual proteoCHIP EVO 96 for SCP 4 proteoCHIP EVO 96 – cellenONE Runs for Figure 1. proteoCHIP EVO 96 inside the cellenONE holder Place the proteoCHIP(s)_EVO_96 inside the cellenONE holder(s) (ref. C-PEVO-96-CHB) and insert the holder(s) containing the proteoCHIP inside the instrument on top of the cooling plates (see Figure 1).
User Manual proteoCHIP EVO 96 for SCP 4.1 Run 1: Master Mix dispensing Prepare 200 µL of Master Mix: 20 µL 1M TEAB + 120 µL H O + 40 µL 1% DDM + 20 µL 100 ng/µL Trypsin (or Trypsin/LysC). Do not vortex the sample, but gently pipette the solution up and down several times to avoid bubbles.
User Manual proteoCHIP EVO 96 for SCP 4.2 Run 2: Cell Isolation Prepare your cell suspension as described in Paragraph 2.3. Main tab • Probe: “cellenWASH” • Run: “EVO_96_SCP_Run_2_Cell_Isolation” or “EVO_96_SCP_Run_2_Cell_Isolation_NoUptake” • Target: “proteoCHIP_EVO_96” Nozzle Setup tab • For dispensing the Cell suspension decrease the Voltage back to its original value and press “Set Nozzle Parameters”.
User Manual proteoCHIP EVO 96 for SCP 4.3 Run 3: Incubation at 50 °C (1h) Main tab • Probe: “cellenWASH” • Run: “EVO_96_SCP_Run_3_Incubation_50C” • Target: “proteoCHIP_EVO_96” Nozzle Setup tab • Increase the Frequency from 500 Hz to 1000 Hz when processing two proteoCHIPs EVO 96 in parallel, and press “Set Nozzle Parameters”.
User Manual proteoCHIP EVO 96 for SCP 4.4 Run 4: Cooldown to 20 °C (~30min) Main tab • Probe: “cellenWASH” • Run: “EVO_96_SCP_Run_4_Cooldown_20C” • Target: “proteoCHIP_EVO_96” Nozzle Setup tab • Keep all the settings from the previous run. Target Setup tab •...
User Manual proteoCHIP EVO 96 for SCP 4.5 Run 5: Dilution For dilution use 0.1% TFA or 0.1% FA (Mass Spec grade). In the end 3.2 µL of solution is added to each well resulting in a total volume of ~3.5 µL. Main tab •...
User Manual proteoCHIP EVO 96 for SCP 4.6 Sample transfer 4.6.1 Option 1 – Transfer to Evotips Figure 2: proteoCHIP EVO 96 inside of the corresponding Holder. The sharp corner of the chip is shown in the bottom right corner. (A) Cell sorting order within a single cell isolation Run.
User Manual proteoCHIP EVO 96 for SCP • Since only ~3 µL of sample is transferred, preload the Evotips with only 17 µL Solvent A instead of 20 µL at the correct Evotip protocol step (“Loading”). • Gently place the proteoCHIP EVO 96 with the diluted samples turning it upside down on top of the Evotips and close the Evotip box lid.
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