Mar 07, 2022

Public workspacePEI Coating

This protocol is a draft, published without a DOI.
  • 1Insitro
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Protocol Citation: kat.titterton 2022. PEI Coating. protocols.io https://protocols.io/view/pei-coating-b5y9q7z6
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it’s working
Created: March 07, 2022
Last Modified: March 07, 2022
Protocol Integer ID: 59137
Abstract
PEI coating 96 and /or 384 wps for highly adherent NGN2 iN replating on DIV3

Based on SOP from Max+ Bobby / Background optimization results

Also refer to: https://www.protocols.io/view/pei-laminin-coating-b5y8q7zw
Protocol materials
ReagentPolyethylenimine (branched)Merck MilliporeSigma (Sigma-Aldrich)Catalog #408727-100ML
Step 1
ReagentGibco™ Distilled WaterGibco - Thermo FischerCatalog #15-230-170
In 3 steps
PEI Stock Prep (1x)
PEI Stock Prep (1x)
Add Amount500 mg of ReagentPolyethylenimine (branched)Sigma AldrichCatalog #408727-100ML to a 50mL conical tube.
Note
PEI stock is an extremely high viscosity liquid. To weigh, Torr a liquid vessel of appropriate volume and use an object to 'stick' goopy PEI on the side to weigh.



Add Amount30 mL of ReagentGibco™ Distilled WaterGibco - Thermo FischerCatalog #15-230-170 to the tube

Heat the PEI solution t0 ~Temperature37 °C on beads. Mix intermittently until PEI is dissolved.
Note
PEI takes a while to go into solution: Shake conical and/or bottle vigorously ever 5-10 min. Takes at least 30 min to fully into solution. Can use hot water directly from tap as a bath to speed up heat transfer.


Add ReagentGibco™ Distilled WaterGibco - Thermo FischerCatalog #15-230-170 to bring volume up to Amount45 mL . Mix again.

Filter-sterilize: pour concentrated PEI solution into a 500mL filter flask.

FOR 1x stock final [PEI] = Amount1 mg /Amount1 mL : Add ReagentGibco™ Distilled WaterGibco - Thermo FischerCatalog #15-230-170 to bring volume up to 500mL.

Aliquot PEI solution into 50mL conicals. Use immediately or proceed to step 7

Store in Temperature-20 °C freezer #13 shelf #9