Lipopolysaccharide-free FH Protein Injection

All materials administered to animals were subjected to lipopolysaccharide removal,^{44 (link)} and were confirmed to be lipopolysaccharide free by the method of Moesby et al.^{45 (link)} Based on the FH dose used successfully in Fakhouri et al.,^{14 (link)} mice were injected intraperitoneally with serum-derived full-length human FH (3 nmol/465 μg per animal), FH^{1–5^18–20} (12 nmol/710 μg), or FH^1–5 (12 nmol/424 μg) in identical volumes of PBS, or PBS alone. Blood was collected onto EDTA via tail venesection before injection and at serial time points thereafter, with plasma separated via centrifugation for storage at −80 °C. Mice were killed at the indicated time points, and the kidneys were collected into PBS and snap frozen in OCT embedding matrix (CellPath, Newtown, UK) for storage at −80 °C.

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Nichols E.M., Barbour T.D., Pappworth I.Y., Wong E.K., Palmer J.M., Sheerin N.S., Pickering M.C, & Marchbank K.J. (2015). An extended mini-complement factor H molecule ameliorates experimental C3 glomerulopathy. Kidney International, 88(6), 1314-1322.

Publication 2015

OCT embedding matrix

Animal Blood Centrifugation Edta Frozen Human Kidneys Lipopolysaccharide Mice Plasma Serum Tail Venesection

Corresponding Organization : Newcastle University

Other organizations : Centre for Inflammation Research, Imperial College London

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Variable analysis

independent variables

Serum-derived full-length human FH (3 nmol/465 μg per animal)
FH^1–5^18–20 (12 nmol/710 μg)
FH^1–5 (12 nmol/424 μg)

dependent variables

Blood collected onto EDTA via tail venesection before injection and at serial time points thereafter, with plasma separated via centrifugation for storage at −80 °C
Kidneys collected into PBS and snap frozen in OCT embedding matrix (CellPath, Newtown, UK) for storage at −80 °C

control variables

PBS alone

positive controls

Serum-derived full-length human FH (3 nmol/465 μg per animal)

negative controls

PBS alone

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