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MB Sample ID: SA171983
Local Sample ID: | S31 |
Subject ID: | SU001925 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Genotype Strain: | APPSwe/PS1ΔE9 (APP/PSEN1) |
Age Or Age Range: | 12 month |
Gender: | Male and female |
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Sample Preparation:
Sampleprep ID: | SP001931 |
Sampleprep Summary: | For liver samples, frozen tissues were first individually pulverized using a mortar and pestle followed by lysing in 1 mL ice-cold lysis buffer ((1:1:2, ACN:MeOH:ammonium bicarbonate (0.1 M, pH 8.0) (LC-MS grade)). Individual samples were sonicated using a probe tip sonicator, 10 pulses, at 30% power, and cooled on ice between samples. A bicinchoninic acid (BCA) protein assay was used to determine the protein concentration for each individual sample and adjusted to a total amount of protein of 200 µg. For untargeted MS studies, SIL-ISs biotin-D2 and phenylalanine-D8 were added to each sample to assess sample processing steps (metabolite extraction and reconstitution). Following lysis and addition of SIL-ISs, protein precipitation was performed by adding 800 µL of ice-cold methanol (4x by volume). Samples were incubated at -80°C overnight. Following incubation, samples were centrifuged at 10,000 rpm for 10 min. The supernatant containing metabolites were dried in vacuo and metabolite extracts were stored frozen at -80 °C until ready to use. |
Processing Storage Conditions: | -80℃ |
Extraction Method: | Following lysis and standard addition, protein precipitation was performed by adding 800µL of ice-cold methanol (4x by volume). Samples were incubated at -80°C overnight. Following incubation, samples were centrifuged at 10,000 rpm for 10 min to eliminate proteins. The supernatants containing metabolites were dried via speed-vacuum. |
Extract Storage: | -80℃ |