PTPSP
Polyacrylamide gel electrophoresis (SDS-PAGE)
français | english
Navigation
Home
Sitemap
This wiki
This page

Polyacrylamide gel electrophoresis (SDS-PAGE), written by Rosa Schier

MES SDS Running Buffer: dissolve one powder bag in 1L of ddH2O (from GenScript)

  • To prepare the final sample of a purification (we want 4ug of protein in 20ul of sample = 4ug protein + …H2O + 10ul loading buffer (calculations with concentration))
  • Heat up the samples 5min at 95°C
  • Put the gel (from Invitrogen) in the tank (be careful to take off the white sticker and take out the comb)
  • Put MES SDS Running Buffer and rinse the holes
  • Put 10ul of sample in each hole and 5ul of AcuteBand Pre-stained Protein Ladder (from lubio science) (don’t put the ladder in the middle)
  • Let it run for 30min at 180V and 400mA + write down the order of the samples
  • Take out the gel, put it in QuickBlue Protein Stain (from lubio science) and let it shake slowly
  • Rinse the tank with water
  • Once the gel is stained rinse with water
  • Scan the gel

 

 

Search
Share