LC3B Antibody Kit for Autophagy

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	Primary antibody 1:1000 dilution

Protocol tips
Primary antibody 1:1000 dilution

Publication protocol

The antibodies were purchased from Millipore unless otherwise stated. Antibodies used are as follows: anti-calbindin (1:1000), anti-calretinin (1:1000), anti-ChAT (1:500), anti-Crym (1:500; Sigma), anti-Ctip2 (1: 500, Abcam), anti-Err3 (1:500, R&D Systems), anti-GFP (1:1000, Invitrogen; or 1:1000, Abcam), anti-LC3B (1:1000, Invitrogen), anti-Satb2 (1:1000), and anti-UCHL1 (1:1000). Antigen retrieval was performed for anti-Ctip2 and anti-Err3 immunocytochemistry (Zhao et al., 2007). Briefly, sections were treated with 5 mM dithiothreitol and 0.1% SDS in Tris-buffered saline for 30 min followed by washes with PBS and incubation in 0.01 M sodium citrate, pH 9.0, at 75°C water bath for 3 h before application of blocking solution (PBS, 0.05% BSA, 2% FBS, 1% Triton X-100, and 0.1% saponin) and incubation with primary antibody solution overnight at 4°C. Appropriate secondary fluorescent antibodies (1:1000, AlexaFluor-488-conjugated, Invitrogen; FITC-conjugated, Invitrogen; or Cy3-conjugated) were added to blocking solution at room temperature for 2 h in the dark. AlexaFluor-555-conjugated α-bungarotoxin (1:500; Invitrogen) was included in the primary antibody solution to label NMJ when appropriate.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for LC3B Antibody Kit for Autophagy below.

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