Fetal Cytokine Profiling Protocol

Arterial blood was collected via the fetal carotid artery catheter before injurious ventilation (Time = 0), at the end of injurious ventilation (15 min), and post-surgery until delivery (1, 3, 6, 12, 24 h) for assessment of plasma proteins (IL-6 and IL-8) using a sandwich enzyme-linked immunosorbent assay (ELISA) assay as described previously (Kelly et al., 2021 (link)). Plates were read on a SpectraMax i3 microplate reader (Molecular Devices, CA, United States) at 450 nm to determine optical density (OD). Standards (recombinant ovine IL-6 or IL-8; Kingfisher Biotech, MN, United States) were included and a standard curve was generated for every ELISA plate used (R² > 0.99).

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Chan K.Y., Tran N.T., Papagianis P.C., Zahra V.A., Nitsos I., Moxham A.M., LaRosa D.A., McDonald C., Miller S.L., Galinsky R., Alahmari D.M., Stojanovska V, & Polglase G.R. (2022). Investigating Pathways of Ventilation Induced Brain Injury on Cerebral White Matter Inflammation and Injury After 24 h in Preterm Lambs. Frontiers in Physiology, 13, 904144.

Publication 2022

SpectraMax i3 microplate reader

Arterial blood Carotid artery Catheter Delivery Devices Enzyme linked immunosorbent assay Fetal Optical Ovine Plasma proteins Surgery

Corresponding Organization : Monash University

Other organizations : King Saud Medical City

Top 5 similar protocols

Variable analysis

independent variables

Injurious ventilation

dependent variables

Plasma proteins (IL-6 and IL-8)

control variables

Arterial blood collected via the fetal carotid artery catheter
Time points: 0 min, 15 min, 1 h, 3 h, 6 h, 12 h, 24 h
Sandwich enzyme-linked immunosorbent assay (ELISA) used to assess plasma proteins
Optical density (OD) measured at 450 nm
Standards (recombinant ovine IL-6 or IL-8) included, with standard curve generated for every ELISA plate (R^2 > 0.99)

controls

Positive control: Recombinant ovine IL-6 or IL-8 standards
Negative control: Not explicitly mentioned

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