Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA098174

Local Sample ID:	Pooled_CKD_Male
Subject ID:	SU001427
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Genotype Strain:	C57BL6J
Age Or Age Range:	18-20 weeks
Weight Or Weight Range:	20-30g
Gender:	Male and female
Animal Animal Supplier:	Jackson Laboratories
Animal Housing:	5/cage
Animal Light Cycle:	12h
Animal Feed:	Ad libitum. Control mice received custom casein-diet. Chronic kidney disease was induced by supplementing casein-based diet with 0.15% adenine
Animal Water:	Ad libitum

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Sample Preparation:

Sampleprep ID:	SP001435
Sampleprep Summary:	Muscles were thawed on ice, weighed, and homogenized with a Teflon-tipped conical pestle with a metal rod (Micro-Tube Sample Pestle with Conical Teflon Tip, fits 1.5ml Tubes, autoclavable at 121°F; Research Products International Corp; 199221; Fisher Scientific). The pestle was rinsed with 2-Propanol, water, and methanol and patted dry with a KimWipe in between samples. The samples were centrifuged to pellet the tissue debris and protein concentrations were quantified on the QuBit. The samples were normalized to 500µg/mL of protein with 5mM Ammonium Acetate in water prior to extraction for a total volume of 100µL. 25µL of sample was aliquoted into a clean tube and extracted with 5µL of Global Metabolomics IS and 200µL of 8:1:1 Acetonitrile:Methanol:Acetone to precipitate proteins. The samples were incubated at 4°C for 30 min and centrifuged at 20,000xg at 4°C for 10min. 200µL of supernatant was transferred to a clean Eppendorf tube and dried under nitrogen gas at 30°C and then reconstituted at 25µL in Global Metabolomics Inj. Std. Mix.

Sampleprep ID:

SP001435

Sampleprep Summary:

Muscles were thawed on ice, weighed, and homogenized with a Teflon-tipped conical pestle with a metal rod (Micro-Tube Sample Pestle with Conical Teflon Tip, fits 1.5ml Tubes, autoclavable at 121°F; Research Products International Corp; 199221; Fisher Scientific). The pestle was rinsed with 2-Propanol, water, and methanol and patted dry with a KimWipe in between samples. The samples were centrifuged to pellet the tissue debris and protein concentrations were quantified on the QuBit. The samples were normalized to 500µg/mL of protein with 5mM Ammonium Acetate in water prior to extraction for a total volume of 100µL. 25µL of sample was aliquoted into a clean tube and extracted with 5µL of Global Metabolomics IS and 200µL of 8:1:1 Acetonitrile:Methanol:Acetone to precipitate proteins. The samples were incubated at 4°C for 30 min and centrifuged at 20,000xg at 4°C for 10min. 200µL of supernatant was transferred to a clean Eppendorf tube and dried under nitrogen gas at 30°C and then reconstituted at 25µL in Global Metabolomics Inj. Std. Mix.