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Table 3 Characteristics of primary antibodies and staining details

From: Neuroinflammation is increased in the parietal cortex of atypical Alzheimer’s disease

Antibody

Antigen

Species

Origin details

Dilution

Incubation time

Antigen retrieval

Detection method

pTau, clone AT8

Tau phosphorylated at Ser202 and Thr205

Mouse IgG1

ThermoFisher, Pittsburgh, USA

1:10000

32 min at 36 °C

Heat-induced (pH 8.5) for 24 min

Optiview

Amyloid-beta, clone IC-16

N-terminus of amyloid-beta (aa 1-16)

Mouse IgG2a

Dr. Carsten Korth, University of Dusseldorf, Germany

1:25

32 min at 36 °C

Heat-induced (pH 8.5) for 24 min

Optiview

GFAP, clone EP672Y

Glial fibrillary acidic protein

Mouse

Roche, Basel, Switzerland

1: 2

8 min at 37 °C

Heat-induced (pH 8.5) for 32 min

Optiview

Iba1

C-terminus of Iba1

Rabbit

Wako Pure Chemical Industries, Osaka, Japan

1:4000

32 min at 36 °C

Heat-induced (pH 8.5) for 16 min

Optiview

CD68, clone KP1

CD68

Mouse IgG1

Dako, Glostrup, Denmark

1:1200

Overnight at 4°C

Heat-induced (pH 6.0) by autoclave

EnVision

HLA-DP/DQ/DR, clone CR3/43

Alpha and beta-chains of all products of the DP, DQ, and DR subregions

Mouse IgG1

Dako

1:800

Overnight at 4°C

Heat-induced (pH 6.0) by autoclave

EnVision

C1q

C1q

Rabbit

Dako

1:25600

Overnight at 4°C

Heat-induced (pH 6.0) by autoclave

EnVision

C3d

C3d

Rabbit

Dako

1:3200

Overnight at 4°C

Heat-induced (pH 6.0) by autoclave

EnVision

C4b

C4b

Rabbit

Abcam, Cambridge, United Kingdom

1:1600

Overnight at 4°C

Heat-induced (pH 6.0) by autoclave

EnVision

C5b-9, clone WU13-15

Neoepitope on C9 in the membrane attack complex

Mouse

Hycult Biotech, Plymouth meeting, USA

1:400

Overnight at 4°C

Heat-induced (pH 6.0) by autoclave

EnVision