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Table 2 Immunoperoxidase methods used by the central laboratory (AGC/FL) and the three testing laboratories

From: Evaluation of alpha-synuclein immunohistochemical methods for the detection of Lewy-type synucleinopathy in gastrointestinal biopsies

 

Primary antibody

Epitope exposure

Signal development

AGC/FL (Angers, France)

p-synuclein, #64

WAKO, Osaka

1:30,000

Formic acid (80 %)

20 min

BOND MAX

Leica Biosystems, Newcastle Upon Tyne, U.K.

TB (Sun City, AZ)

p-synuclein, polyclonal

H. Akiyama, Tokyo

1:10,000

Proteinase K 1:100

37 ° C, 20 min

ABC, DAB

Vector Laboratories

JK (Chicago, IL)

Alpha-synuclein, LB509

Invitrogen

1:500

Formic acid (88 %)

20 min

ABC, DAB

Vector Laboratories

DM (Toronto, Canada)

p-synuclein, #64

WAKO, Osaka

1:12,000

Ventana Protease 2

72 ° C, 8 min

Citrate buffer, pH 6 4

90 ° C, 4 min

Ventana Benchmark Ultra,

Optiview multimer and amplifier

  1. p-synuclein = alpha-synuclein phosphorylated at serine 129; ABC = avidin-biotin peroxidase complex; DAB = 3, 3’ di-aminobenzidine