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  • Preparation of Purified Gram-positive Bacterial Cell Wall and Detection in Placenta and Fetal Tissues.

Preparation of Purified Gram-positive Bacterial Cell Wall and Detection in Placenta and Fetal Tissues.

Bio-protocol (2017-06-03)
Beth Mann, Lip Nam Loh, Geli Gao, Elaine Tuomanen
摘要

Cell wall is a complex biopolymer on the surface of all Gram-positive bacteria. During infection, cell wall is recognized by the innate immune receptor Toll-like receptor 2 causing intense inflammation and tissue damage. In animal models, cell wall traffics from the blood stream to many organs in the body, including brain, heart, placenta and fetus. This protocol describes how to prepare purified cell wall from Streptococcus pneumoniae, detect its distribution in animal tissues, and study the tissue response using the placenta and fetal brain as examples.

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Sigma-Aldrich
磷酸钾二元 二元, ACS reagent, ≥98%
Sigma-Aldrich
乙二胺四乙酸 二钠盐 二水合物, suitable for electrophoresis, for molecular biology, 99.0-101.0% (titration)
Sigma-Aldrich
磷酸钾 一元, powder, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99.0%
Sigma-Aldrich
L-谷氨酰胺, meets USP testing specifications, suitable for cell culture, 99.0-101.0%, from non-animal source
Sigma-Aldrich
烟酸, ≥98%
Sigma-Aldrich
氯化锂, for molecular biology, ≥99%
Sigma-Aldrich
(−)-核黄素, from Eremothecium ashbyii, ≥98%
Sigma-Aldrich
L-天冬酰胺 一水合物, BioReagent, suitable for cell culture