Okay, here’s a breakdown of the methods described in the provided text, organized for clarity. I’ll categorize it into sections: Study Approval/Ethics, ELISA (NOTCH3-ECD Quantification), Cross-Reactivity Testing, Immunoprecipitation & Western Blotting, and Statistical Analysis.
1. Study Approval/Ethics
* The study received approval from multiple Institutional Review Boards (IRBs):
* university of Arizona (protocol no. 1100000621A013)
* University of Alabama (protocol no. 1639383-7)
* University of New Mexico (protocol no. 00003255)
* University of California, Los Angeles (UCLA) (protocol no. 12-0738)
* Americas Hospital, Guadalajara, Mexico (protocol no. AH-IRB-B2)
* mount Sinai, New York (protocol no.RC-4590)
* University of Cambridge (UK) – part of the UK National Cohort Study of Idiopathic and Heritable Pulmonary Arterial hypertension (ClinicalTrials.gov ID: NCT01907295; UK Research ethics Committee reference no. 13/EE/0203)
* Informed consent was obtained as part of the UK National Cohort Study.
2. ELISA (NOTCH3-ECD Quantification)
* Purpose: To measure levels of NOTCH3-ECD in human serum.
* Kit: Human NOTCH3-ECD ELISA kit (Cloud Clone, cat.no. SEL147Hu)
* Blinding: Operators were blinded to the case/control status of samples.
* Standard Curve:
* High standard: 100 ng/ml (reconstituted from standard diluent)
* Serial dilutions: 10
