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85 lines
2.5 KiB
YAML
85 lines
2.5 KiB
YAML
---
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# Protocol metadata
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id: PROT-0013
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name: BSA-OAPA and Insulin Combined Treatment Protocol
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version: 1.0
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description: Protocol for treating cells with BSA-conjugated oleic/palmitic acids and insulin
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author: JJ
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created: 2025-04-30
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last_updated: 2025-05-07
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category: cell-treatment
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# Treatment details
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treatment:
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primary:
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type: fatty-acid
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ratio: "1:1 (OA:PA)"
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total_concentration: 0.5 mM
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duration: 24 hours
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secondary:
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type: hormone
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name: Insulin
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concentration: 100 nM
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duration: 24 hours
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control:
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- BSA-only treatment
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- Vehicle (no insulin)
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# Materials required
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materials:
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- name: Oleic acid (OA)
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concentration: 0.25 mM
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preparation: BSA-conjugated
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- name: Palmitic acid (PA)
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concentration: 0.25 mM
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preparation: BSA-conjugated
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- name: Insulin solution
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concentration: 100 nM
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preparation: Diluted in appropriate buffer
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- name: Complete culture medium
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notes: Use media appropriate for cell type
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# Protocol steps
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steps:
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- step: 1
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action: "Prepare BSA-conjugated OA and PA mixture"
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details: "Final concentration: 0.25 mM OA, 0.25 mM PA (0.5 mM total, 1:1 ratio)"
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- step: 2
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action: "Prepare insulin working solution"
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details: "Dilute stock insulin to 100 nM final concentration"
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- step: 3
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action: "Treat cells with OAPA and insulin simultaneously"
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details: "Add both treatments to culture medium before applying to cells"
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- step: 4
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action: "Incubate for 24h with combined treatment"
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details: "Maintain standard culture conditions during incubation"
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- step: 5
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action: "Set up appropriate controls"
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details: "Include BSA-only control, insulin-only control, and untreated control"
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# Experimental context
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experimental_notes:
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- condition: "Combined fatty acid and insulin stimulation"
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- condition: "Medium dose OAPA (0.5 mM total)"
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- condition: "Standard insulin dose (100 nM)"
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- note: "This protocol tests the interaction between fatty acid loading and insulin signaling"
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# Safety considerations
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safety:
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ppe: "Lab coat, gloves and eye protection required"
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hazards: "Avoid direct contact with concentrated fatty acids"
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# Data collection
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data_collection:
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timepoints: [0h, 6h, 12h, 24h]
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measurements:
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- "Insulin signaling pathway activation (Western blot)"
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- "Lipid accumulation (Oil Red O staining)"
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- "Gene expression changes (qPCR)"
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# References
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references:
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- "Internal protocol based on standard BSA-fatty acid conjugation methods"
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- "Liu Y, et al. (2022) Effects of insulin and fatty acids on metabolic signaling. J Cell Biol 35(2):122-134"
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---
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