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Merck

Platelet Migration and Bacterial Trapping Assay under Flow.

Bio-protocol (2018-09-20)
Shuxia Fan, Michael Lorenz, Steffen Massberg, Florian Gaertner
ABSTRACT

Blood platelets are critical for hemostasis and thrombosis, but also play diverse roles during immune responses. We have recently reported that platelets migrate at sites of infection in vitro and in vivo. Importantly, platelets use their ability to migrate to collect and bundle fibrin (ogen)-bound bacteria accomplishing efficient intravascular bacterial trapping. Here, we describe a method that allows analyzing platelet migration in vitro, focusing on their ability to collect bacteria and trap bacteria under flow.

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Sigma-Aldrich
Tampone fosfato salino di Dulbecco, Modified, without calcium chloride and magnesium chloride, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Dimetil solfossido, sterile-filtered, BioPerformance Certified, meets EP, USP testing specifications, suitable for hybridoma
Sigma-Aldrich
HEPES, 1 M, pH 7.0-7.6, sterile-filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
Brodo LB (Lennox), Highly-referenced microbial growth powder medium, low salt, suitable for salt-sensitive E.coli culture.
Sigma-Aldrich
Ampicillina
Sigma-Aldrich
Trombina, lyophilized powder, 40-500 NIH units/mg protein (biuret)
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Adenosine 5′-diphosphate sodium salt, bacterial, ≥95% (HPLC)
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Fibrinogeno, 50-70% protein (≥80% of protein is clottable)
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Sodio citrato tribasico, ACS reagent, ≥99.0%
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Albumina, Cellastim S, recombinant, expressed in rice, lyophilized powder, suitable for cell culture, low endotoxin, ≥96% (SDS-PAGE)
Sigma-Aldrich
IPTG, ≥99% (TLC), ≤0.1% Dioxane