WB

title: WB updated: 2024-02-13T12:30:26 created: 2023-11-27T10:46:26

• Lysates

• BCA kit

  • WR = 50:1 reagent A:B

• 10 ul standard

• 10 usually 10x dil lysate

• Smpls

  • Laemmli
  • Dil w RIPA
  • Aim for ca 20-30 ug protein/20ul

• EL FO

  • Dddddddd
  • Fit thevgel to the thing - no leak etc, right directiom
    • Fake gel rightdirection
  • 80-140 V, start lower

• Transfer -

  • PAPER thing 10layer 2x dil soak in trans vuffer
  • Membrane MeOH 1 min +
  • Sandwich
    • Right side and all
  • 25 v 10-20 min

• Block milk + tbst 1 h rt

• Primary in bsa 1% in tbst overnight 4c

• Wash 3x 10 min

• Secondary 1 ul in 20ml 1%bsa in tbst

• 1 h in dark. Rt

• Wash 3x 10 min

• Cca 1 ml of each ECL reagent

• Dadada

  • Best to capture 10 images for 600 s