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Western Blot Protocol Diagram

By FigPad · Updated Apr 22, 2026

A ten-step western blot workflow figure for methods sections, lab protocols and teaching slides, from protein extraction to ECL imaging.

Step-by-step western blot protocol diagram showing protein extraction, BCA assay, SDS-PAGE, semi-dry transfer, Ponceau S staining, blocking, antibody incubation and ECL detection

What this figure shows

  • Protein extraction in RIPA buffer with protease and phosphatase inhibitors
  • BCA protein quantification on a 96-well plate with a standard curve
  • Sample denaturation in SDS loading buffer at 95 °C for 5 min
  • SDS-PAGE on a 4% stacking and 10% resolving gel with a protein ladder
  • Semi-dry transfer to a PVDF membrane (25 V, 30 min) and a Ponceau S check
  • Blocking in 5% BSA/TBST, primary antibody at 4 °C overnight, HRP secondary for 1 h
  • ECL detection with example target and GAPDH loading-control bands

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