Sample preparation

Sample preparation instructions

SAMPLE PREPARATION | Shipping


Whole blood (human) 

-Blood should be collected into the smallest EDTA vacutainer (2 ml)  
-IMPORTANT! If possible, the blood sample could be divided into 500 µl sample aliquots in separate tubes, e.g. Eppendorf tube 
-Samples should be frozen at -20 °C as soon as possible after withdrawal for several days. Samples should be stored for longer time in -80 °C 
IMPORTANT! Keep consistent time between withdrawal and freezing for all samples   

Whole blood (animal) 
-Blood should be collected into the EDTA tubes and divided into 200 µl sample aliquots in separate tubes, e.g. Eppendorf tube 
-Samples should be frozen at -20°C as soon as possible after withdrawal for several days.
-Samples should be stored for longer time in -80°C 

IMPORTANT! Keep consistent time between withdrawal and freezing for all samples   

PLEASE NOTE:
-2 ml/500 µl (human) or 200 µl (animal) of whole blood is enough for the measurement of all four NAD metabolites 
-Plasma or serum are not suitable for NADmed analysis 
-NAD levels are normalized per volume (final concentration is in µM) .
-Optional: NAD levels can be normalized per protein amount .

Tissues (human/animal)  
IMPORTANT: to reduce variability between the samples from different subjects/animals, it is very important to take aliquots of tissue from exactly same areas of the organ .

Fresh tissue samples:
-Organ/tissue sample should be collected by standard method, rinsed with cold PBS and the excess of buffer removed with paper towel 
-Each organ/tissue sample should be 10-20 mg, the exact weight of each sample piece should be recorded  Samples should be snap frozen in liquid nitrogen and stored in -80°C.   

Frozen tissue samples:  
If samples need to be aliquoted, it should be done under liquid nitrogen to avoid sample melting  The weight of each frozen sample should be recorded   Samples should be stored in -80 C   
PLEASE NOTE:  NAD levels are normalized per sample weight  
Optional: NAD levels can be normalized per protein amount   

Cultured cells
-One 10 cm plate (confluency 90-100 %) is suitable for the measurement of all four NAD metabolites 
-Cells should be grown in 10 cm plates until 90-100 % confluency, then washed with excess of PBS 
-Cells should be collected by scraping in PBS (not trypsin) and centrifuged (750 rpm) 
-After removing the supernatant, snap frozen in liquid nitrogen and stored at -80 C 
-NAD levels are normalized per protein amount   

Pseudonymisation
All samples should be pseudonymized and labelled only with sample-specific code. We also recommend to randomize the order of the samples.   Basic information for each sample required in a separate sheet  (Sample code,  Sample source (e.g. human) , Sample type  (e.g. whole blood) and volume  or  weight of each sample) .

Shipment  Samples should be shipped on dry ice. The amount should be sufficient enough to keep the sample frozen for several days. The institute is open on week days 7-19.   

Shipping address:  Liliya Euro/Wartiovaara lab  Biomedicum 1  Haartmaninkatu 8  00290 Helsinki  Finland   

Contact: Liliya Euro  (liliya.euro@helsinki.fi)      

+358406811716
Vuorikatu 7, 00100 Helsinki, Finland