PQ dichloride (1,1-dimethyl-4,4-dipyridinium dichloride) and MTX were purchased from FUJIFILM Wako Pure Chemical Corporation (Osaka, Japan). Rabbit antibodies against ACSL4 (FACL4: ab155282) were purchased from Abcam (Cambridge, MA, USA). Mouse monoclonal antibodies against β-actin were purchased from Sigma Aldrich (St. Louis, MO, USA). Rat monoclonal antibodies against mouse Ly6G (clone 1A8) were purchased from Bio X cell (Lebanon, NH, USA). The DAKO Envision+ system peroxidase kit was purchased from DAKO (Carpinteria, CA, USA). Anti-mouse CD11b-FITC (11-0112-82), anti-mouse Ly-6G/6C-PE (12-5931-82), and anti-mouse F4/80-PE-Cy5 (15-4801-82) antibodies were purchased from eBioscience (San Diego, CA, USA). Ferrostatin-1 (Fer-1) was purchased from Cayman Chemical (Ann Arbor, MI, USA).
Dako envision system peroxidase kit
Manufactured by Agilent Technologies
Sourced in United States, Denmark
The DAKO Envision+ system peroxidase kit is a laboratory equipment product designed for immunohistochemistry (IHC) applications. The kit provides a secondary antibody and a chromogenic substrate system for the detection of primary antibodies bound to target antigens in tissue sections or cell preparations.
Automatically generated - may contain errors
2 protocols using dako envision system peroxidase kit
1
Antibody and Reagent Procurement for Cell Analysis
2
Immunostaining for Cytoskeletal and Extracellular Matrix Proteins
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Immunostaining was performed using a commercially available DAKO EnVision™ System, Peroxidase kit (Dako, Glostrup, Denmark). Primary antibodies used to detect smooth muscle actin (SMA), type 1 collagen, and type 3 collagen were anti-SMA (ab5694, Abcam, Tokyo, Japan; 1:400), anti-collagen I (ab88147, Abcam, 1:100), and anti-collagen III (ab7778, Abcam, 1:1000), respectively. Heat-mediated antigen retrieval (95°C, 20 min) was performed using a buffer balanced to either pH 6.0 (anti-SMA, anti-collagen III) or pH 9.0 (anti-collagen I) before staining. Next, the sections were washed three times with phosphate-buffered saline (PBS) and incubated with the primary antibody for 30 min at room temperature. Next, the sections were rewashed three times with PBS and incubated with the labeled polymer for 30 min at room temperature. The sections were washed three times in tris-buffered saline (pH 7.4) and incubated with 3,3′-diaminobenzidine as substrate chromogen for 10 min to reveal brown-colored precipitate at the antigen site.
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