Cells were grown on 12 mm coverslips and fixed with 3.6% formaldehyde in PBS for 15 minutes. Cells were permeabilized with PBS containing 0.1% Triton X-100, blocked with 2% BSA in PBS for 30 minutes at RT, and incubated with the appropriate antibodies overnight at 4 °C. PLA was performed using the Duolink anti-Mouse MINUS and anti-Rabbit PLUS In Situ PLA probes (Sigma) and the Duolink In Situ Detection Reagents Red (Sigma), according to the to manufacturer’s instructions. Digital images were acquired using an LSM700 microscope equipped with a 40x Plan-apochromat 1.3 NA oil immersion lens (Carl Zeiss) and nuclear foci were quantified using an ImageJ macro.
Duolink anti mouse minus and anti rabbit plus in situ pla probes
Manufactured by Merck Group
The Duolink anti-Mouse MINUS and anti-Rabbit PLUS In Situ PLA probes are lab equipment used for proximity ligation assay (PLA) analysis. The probes are designed to detect and visualize protein-protein interactions within cells or tissues.
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Lab products found in correlation
2 protocols using duolink anti mouse minus and anti rabbit plus in situ pla probes
1
Proximity Ligation Assay for Protein Interactions
2
Muscle Protein Interaction Analysis
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Mouse skeletal muscle was solubilized as described above. Immunoprecipitations were then performed following the manufacturer’s instructions using the Dynabeads Protein G (Life Technologies). PLAs were performed following the manufacturer’s instructions using the Duolink anti–Mouse MINUS and anti–Rabbit PLUS In Situ PLA probes and the Duolink In Situ Detection Reagents Red (Sigma). Cells were counterstained with α-BTX–A488 for 60 min and imaged using a Zeiss LSM880 microscope with an AiryScan1 detector equipped with a 63× 1.4-NA.
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