Mouse anti cd68 clone pg m1

Formalin-fixed, paraffin-embedded brain and spinal cord tissue were available from two patients (Patients AM2 and AM4). The paraffin sections were cut at 4 µm thickness, mounted on glass slides and stained with routine haematoxylin and eosin, and Luxol® fast blue/cresyl violet histochemical stains. Sections were examined by immunohistochemistry with the following antibodies: rabbit anti-glial fibrillary acid protein (GFAP) (polyclonal, 1:2500, Dako), mouse anti-phosphorylated neurofilaments (clone SMI31, 1:5000, Sternberg), mouse anti-myelin basic protein (clone SMI94, 1:2000, Sternberg), rabbit anti-ubiquitin (polyclonal, 1:1200, Dako), mouse anti-p62 (3/P62LCK Ligand, 1:100, BD Transduction), mouse anti-α-synuclein (clone KM51, 1:50, Leica/Novocastra), mouse anti-amyloid-β (clone 6F3D, 1:100, Dako), mouse anti-hyperphosphorlyated tau (clone AT8, 1:1200, Innogenetics), mouse anti-TDP43 (clone 2E2-D3, 1:3000, Abova), mouse anti-CD68 (clone PG-M1, 1:100, Dako), mouse anti-CD3 (LN10, 1:100, Leica/Novocastra), mouse anti-CD20 (clone 7D1, 1:200, Dako). Immunohistochemistry was carried out either manually or on a BOND-MAX autostainer (Leica Microsystems) using 3,3′-diaminobenzidine as chromogen. Negative controls were treated identically except that the primary antibody was omitted. Appropriate positive controls were used for all immunohistochemical studies.

Immunohistochemical staining was performed using the Dako autostainer platform (Dako, Denmark) as previously described [24 (link)]. Sections were stained using the following primary antibodies: mouse anti-CD68 (clone PG-M1, 1:100, Dako), mouse anti-CD45 (clone 2B11, 1:200, Dako), rabbit anti-Iba1 (ionized calcium binding adaptor molecule 1, 1:1000, Wako), rabbit anti-GFAP (1:2000, Dako), mouse anti-neurofilament (NF) (phosphorylated and non-phosphorylated NF-heavy chain; clone N52, 1:1000, Sigma-Aldrich), mouse anti-IL-1α (clone 4414, 1:1200, R&D Systems), mouse anti-IL-1β (clone 2E8, 1:50, BioRad), rabbit anti-TNF (1:100, ThermoFisher Scientific), rabbit anti-TNFR1 (clone H-271, 1:50, Santa Cruz), rabbit anti-TNFR2 (1:50, Sigma-Aldrich), and rat anti-IL-1Ra (clone 40,007, 1:1500, R&D Systems). The antigen-antibody complex was visualized using EnVision+System horse-radish peroxidase-labelled Polymer (Dako), PowerVision+Poly-HRP IHC (AH Diagnostics), or CSAII (Dako) detection systems.