Selected Laboratory Protocols

Mouse/Atherosclerosis/Metabolism Protocols

• NASH mouse model based on FPC-NASH diet
• Tips on Mouse Handling
• Assay of Hepatic p-CaMKII in Fasting Mice
• Liver Macrophages Isolation
• Genotyping of Asm-/- Mice
• Protocol for Bone Marrow Transplantation
• Principles of Mouse Husbandry: George Kuriakose's 2013 PowerPoint Presentation
• Assessment of Progression of Atherosclerotic Lesions in Mice
• Mouse Shipping Information
• Genotyping of ACAT-/-Mice
• Genotyping of F4/80-Cre Knock-In Mice
• Basic Information for Directing a Mouse Atherosclerosis Project
• Important Information About the Harlan-Teklad 88137 Western Diet Used in the Tabas Laboratory for Mouse Atherosclerosis Studies
• More Detailed Procedure for Embedding Mouse Hearts in OCT in Preparation for Frozen Sections
• Shipment of Mouse Femurs for Use in Isolating Marrow Cells for In Vitro MΦ Studies or BMT
• Fundamental Principles of Conducting Mouse Obesity Studies
• P-Akt, p-HSP27, and p-MK2 Assays in Obese Mice
• DIO vs. ob/ob
• Dex-thymus

Lipoprotein Protocols

• Protocol for Reconstitution of the Hydrophobic Core of Acetyl-LDL
• Labeling of Acetyl-LDL with DiI (1,1'-dioetadeeyl-3,3,3',3'-tetramethylindocarboeyanine perchlorate)
• Protocol for Preparation of LDL from Human Plasma
• Precautions to avoid endotoxin contamination of LDL and Acetyl-LDL

Cell Protocols

• L Cell Conditioned Medium (LCCM)
• Preparation of Adherent Macrophages for Electron Microscopy
• Technical Points About Plasmid-Mediated Transfection or RNAi-Mediated Gene Knockdown in Methyl-BSA-Elicited Macrophages Using the Amaxa Nucleofector® System (Dr. Yankun Li, September, 2005)
• Concanavalin A-Elicited Mouse Peritoneal Macrophages
• Preparation of Peritoneal Macrophages from Methylated BSA-Immunized Mice
• Filipin Fluorescence Staining of Free Cholesterol in Cultured Cells
• Isolation of bone marrow macrophage
• Isolating Cytosolic and Mitochondrial Extracts from Mouse Macrophages
• Isolating Smooth Muscle Cells from Mouse Aorta
• Preparation of Apoptotic Jurkat Cells for Efferocytosis Assays
• Protocol – Flow cytometry analysis of IL-10 and TGF-ß
• PMN isolation protocol
• Seahorse XF96 Protocol – adapted from Agilent
• Human macrophage isolation

Assays

• Standardized Tabas Laboratory In-Vitro Efferocytosis Engulfment Assay
• Immunoprecipitations Using TrueBlot
• GC Assay for Cellular Cholesterol Mass
• CHOP Immunoblots of Whole Cell Extracts
• Preparation of ¹⁴ C-Oleate-Albumin Solution for Whole-Cell Cholesterol Esterification Assay
• Method for Preparing Cytoplasmic and Nuclear Extracts for Immunoblots of UPR Proteins
• Assay for Apoptosis of FC-Loaded Macrophages (Annexin/propidium iodide kit is from Molecular Probes)
• Extraction of Lipids in Solution by the Method of Bligh & Dyer
• Euglobulin Clot Lysis Assay for Mouse Plasma