The human RPE cell line ARPE-19 was purchased from ATCC (Manassas, VA, US). DMEM/F-12, HEPES medium (#11330-057) was purchased from Life Technologies (Grand Island, NY, US). FBS (#SCRR-30-2020) was obtained from ATCC. Penicillin-Streptomycin (#15140122) was obtained from Life Technologies. Recombinant human IL-1α (200-LA-002) was obtained from R&D Systems (Minneapolis, MN, US). Cholesterol crystals (#C8667) were purchased from Sigma-Aldrich (St. Louis, MO, US). Bay 11-7082 inhibitor (#196870) was obtained from EMD Millipore. Anti-IL-18 antibody (#ab137664), anti-IL-6 antibody (#ab32530), and anti-β-actin antibody (ab8227) were obtained from Abcam (Cambridge, MA, US). Anti-IL-1β antibody (#MAB201) and anti-IL-8 antibody (#MAB208) were purchased from R&D Systems. HRP-linked secondary antibodies (7074S, 7076S) were obtained from Cell Signaling Technology (Danvers, MA, US).
Cholesterol crystals
Manufactured by Merck Group
Sourced in United States
Cholesterol crystals are a type of laboratory equipment used for the study and analysis of cholesterol. These crystals provide a visual representation of the molecular structure of cholesterol, which is a naturally occurring lipid found in the body. The core function of cholesterol crystals is to facilitate the investigation and understanding of cholesterol-related processes and conditions.
Automatically generated - may contain errors
Lab products found in correlation
Arpe 19, by American Type Culture Collection (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Anti β actin antibody ab8227, by Abcam (1 mentions)
Anti il 18 antibody ab137664, by Abcam (1 mentions)
Anti il 6 antibody ab32530, by Abcam (1 mentions)
Fetal bovine serum (fbs), by American Type Culture Collection (1 mentions)
Anti il 8, by Cell Signaling Technology (1 mentions)
Hrp linked secondary antibody, by Cell Signaling Technology (1 mentions)
Anti il 8 antibody, by R&D Systems (1 mentions)
Recombinant human il 1α, by R&D Systems (1 mentions)
Anti il 6, by Cell Signaling Technology (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
3 protocols using cholesterol crystals
1
Investigating Inflammatory Responses in ARPE-19 Cells
2
Cholesterol Crystal-Induced Microinfarct Model
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To develop the multiple diffuse microinfarct model, cholesterol crystals (Sigma-Aldrich, Saint Louis, MO, USA) were first filtered through a 70-μm sieve followed by a 40-μm cell strainer, and then resuspended in saline and counted with a hemocytometer. After the animal was anesthetized, we carefully exposed the right common carotid artery (CCA), internal carotid artery (ICA) and external carotid artery (ECA) under a dissecting microscope. The ICA was temporarily ligated, and the distal portion of the ECA was permanently ligated. Subsequently, we clipped the CCA and the proximal parts of the ECA and ICA, and made an incision between the ligation site and the clip site. Then, a polyethylene tube was inserted into the ECA, extending to the CCA. Around 3000 cholesterol crystals in 100 μl of saline or 100 μl saline alone (for sham animals) were injected through the tube after removal of the microvascular clip. The tube was eventually taken out, and the wound was closed after the proximal ECA was permanently ligated. A total of 12 mice out of 48 mice died after induction of multiple diffuse microinfarcts.
3
IL-1α Stimulation of ARPE-19 Cells
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The human RPE cell line ARPE-19 was purchased from ATCC (Cat#CRL-2302, Manassas, VA, US). DMEM/F-12 HEPES medium (Cat# 11330–057), fetal bovine serum (FBS; Cat# 10438–026) and penicillin-streptomycin (100 U/mL-100 μg/ml; Cat# 15140) were obtained from Life Technologies (Grand Island, NY, US). Recombinant human IL-1α was obtained from R&D Systems (Cat# 200-LA-010, Minneapolis, MN, US). Cholesterol crystals were purchased from Sigma-Aldrich (Cat# C8667, St. Louis, MO, US). Anti-IL-6 (Cat# 12153) and anti-IL-8 (Cat#4970) antibody was obtained from Cell Signaling Technology (Danvers, MA, US). anti-IL-8 antibody (Cat# MAB208) was purchased from R&D Systems (Minneapolis, MN, US). Anti-rabbit (Cat# 7074) and anti-mouse (Cat# 7076), HRP-linked secondary antibodies were obtained from Cell Signaling Technology (Danvers, MA, US).
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