Detergent-based Western Blot Stripping Buffer

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100 mL Recipe Desired Volume: mL    
20 mL of 10% SDS
(71736)
  mL of 10% SDS
12.5 mL of 0.5 M Tris-HCl pH 6.8 (FW 157.60)    mL of 0.5 M Tris HCl pH 6.8
0.8 mL of β-mercaptoethanol (63689)   mL of β-mercaptoethanol
Adjust final volume to 100 mL with Milli-Q® Water Adjust final volume to   mL with Milli-Q® Water
  Procedure
 
  1. In a fume hood, place the blot in detergent stripping buffer and incubate with agitation for 30 minutes at 50 °C.
  2. Place the blot in 1X PBS and agitate for 10 minutes. Repeat with fresh buffer.
  3. Optional: Repeat the initial detection protocol (omitting the primary antibody step) to make sure that the antibodies have been inactivated or removed from the membrane.
  4. Place the blot in fresh 1X PBS and agitate for 10 minutes.
  5. Proceed to the blocking step for the next round of immunodetection.

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