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2 protocols using tlr2 clone 6c2

1

Immune Cell Profiling from Brain Tissue

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Brains were isolated and homogenized, and single-cell suspensions were prepared by filtration through a 100 μm mesh. Cellular infiltrates were stained with anti-CD45.2 (clone 104, BD Biosciences), anti-CD11b (clone M1/70, BD Biosciences), anti-Ly-6G/Ly-6C (clone RB6-8C5, BD Biosciences), and/or TLR2 (clone 6C2, eBioscience), and acquired on a FACS Canto (BD Biosciences) running DIVA software.
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2

Dectin-mediated Signaling Pathway Analysis

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Syk inhibitors SykI and BAY 61-3606, JNK inhibitor SP600125, ERK inhibitor U0126, and p38 inhibitor SB203580 were obtained from Calbiochem-Merck. Laminarin and zymosan were purchased from InvivoGen. Peptide inhibitor for NADPH oxidase assembly, gp91ds-tat, was purchased from AnaSpec. Diphenyleneiodonium chloride (DPI) and Mito-TEMPO were obtained from Sigma-Aldrich. LysoTracker Red was obtained from Life Technologies.
Antibodies against LC3, Rubicon, NLRX1, phospho (p)-p40-phox (Thr154), p-JNK (Thr183/Tyr185), p-ERK1/2 (Thr202/Tyr204), p-p38 (Thr180/Tyr182), p-c-Fos (Ser32), p-IKKα/β (Ser176/180), p-IκBα (Ser32), IκBα, and p-NFκBp65 (Ser536) were purchased from Cell Signaling. Antibodies against p-Syk (Tyr525), p-c-Jun (Ser63), and TUFM were obtained from Abcam. Anti-Syk, anti-ATG5, anti-β-actin, HRP-conjugated anti-rabbit IgG, and Rabbit IgG isotype control were purchased from GeneTex Inc. Receptor blocking antibodies against Dectin-1 (clone 2A11), Dectin-2 (clone D2.11E4), and CR3 (clone 5C6) were purchased from Bio-Rad (formerly AbD Serotec) and TLR2 (clone 6C2) was from eBioscience.
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