To detected Fetuin-A/FGF23 interactions in the HepG2 and in the plasma, Duolink (Sigma-Aldrich) in situ experiment was performed following the manufacturer's instructions. Cells were fixed in PFA for 10min, blocked for 30min (Sigma-Aldrich) and stained with anti-FGF23 and anti-Alpha-2-HS-glycoprotein o/n at 4°C. A second incubation with PLUS/MINUS PLA probe was performed and after ligation and amplification (both from Sigma) we visualized the interaction by immunofluorescence. As negative ctrl, we chose to omit one of the two antibodies alternatively.
Plus minus pla probe
Manufactured by Merck Group
The PLUS/MINUS PLA probe is a lab equipment product developed by Merck Group. It is designed to provide accurate and reliable measurements for specific applications. The core function of this probe is to facilitate precise data collection and analysis, without interpretation or extrapolation on its intended use.
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Lab products found in correlation
2 protocols using plus minus pla probe
1
Detecting Fetuin-A/FGF23 Interactions in HepG2 and Plasma
2
Detecting Fetuin-A/FGF23 Interactions via Duolink
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Duolink (Sigma–Aldrich) in situ experiment was performed to detect Fetuin-A/FGF23 interactions in M2-10B4, L88/5, OS mouse cell lines, in the primary ADMSC, BMMSC, CBMSC and in the tibial mouse tissue following the manufacturer’s instructions. Cells and tissue were fixed in PFA for 10 min, blocked for 30 min (Sigma–Aldrich), and stained with anti-FGF23 and anti-Alpha-2-HS-glycoprotein (all antibodies information in Table S2 ) overnight (o/n) at 4 °C. A second incubation with PLUS/MINUS PLA probe was performed and after ligation and amplification (both from Sigma–Aldrich), we visualized the interaction by IF. As control negative, we chose to omit one of the two antibodies alternatively.
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