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Table 3 Western blotting parameters

From: Systemic immune challenges trigger and drive Alzheimer-like neuropathology in mice

Antibody Run time, hours/voltage Transfer time hours Blocking reagent/time, hours Secondary antibody incubation, hours
total Tau/Ab-2 1.5 to 2/125 1.5 10% WBR1/1 1.5
PHF-Tau
pTauT205
APP, C-terminal 2/125 or 2.5/1003 1.5 10% FBS2/2 1
APP (A4) 3/1253 1.5 10% WBR/1 1.5
β-Actin4    10% WBR/1 1
  1. FBS, fetal bovine serum (heat-inactivated); paired helical filament; WBR, Western blocking reagent (Roche Applied Science).
  2. 1Incubation of membrane in boiling PBS for 5 minutes recommended.
  3. 2Incubation of membrane in boiling PBS for 5 minutes required.
  4. 3Western blotting apparatus kept in the water bath at room temperature to prevent overheating.
  5. 4β-actin was measured for each blot after stripping (2% SDS at 56°C for 30 minutes).