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Merck

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
RESUMEN

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
Ácido clorhídrico, ACS reagent, 37%
Sigma-Aldrich
Fosfato de potasio dibasic, ACS reagent, ≥98%
Sigma-Aldrich
Ácido etilenodiaminotetraacético disodium salt dihydrate, suitable for electrophoresis, for molecular biology, 99.0-101.0% (titration)
Sigma-Aldrich
Fosfato de potasio monobasic, powder, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99.0%
Sigma-Aldrich
L-Glutamina, meets USP testing specifications, suitable for cell culture, 99.0-101.0%, from non-animal source
Sigma-Aldrich
Cloruro de litio, for molecular biology, ≥99%
Sigma-Aldrich
(−)-Riboflavina, from Eremothecium ashbyii, ≥98%
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Nicotinic acid, ≥98%
Sigma-Aldrich
L-Asparragina monohydrate, suitable for cell culture, BioReagent