Bio-Rad Human Metabolic and Hormone Assays - Manual (Page 17)

15
Cell culture media
1. Collect cell culture supernatants and centrifuge at 1,000 x g for
15 min at 4°C. For cell lines cultured in serum-free media, collect
samples and add BSA as a carrier protein to a final concentration of
at least 0.5%. This is done to stabilize protein analytes and to prevent
adsorption to labware.
2. Transfer to a clean polypropylene tube. If cellular debris or precipitates
are present, centrifuge again at 10,000 x g for 10 min at 4°C.
3. If high levels of analyte are expected, samples can be diluted in culture
media. Optimal dilution factor must be determined by the end user.
Supplement serum-free media with at least 0.5% BSA final.
4. Assays samples immediately or aliquot and store at –70°C.
Note: Compatibility of a given media type with the assays must be
validated by the end user.
Table 5. Serum and plasma sample dilution guidelines.
Panel Sample
Dilution
Volume of Sample, µl Volume of
Sample
Buffer, µl
Metabolic panel 1 1:5 20 80
Metabolic panel 2 1:5 20 80
Metabolic panel 3 1:500,000 (a) Prepare 1:50 5
(b) Prepare 1:100 5 of (a)
Prepare 1:100 5 of (b) 245
495
495
Metabolic panel 4 1:500 (c) Prepare 1:10 10
Prepare 1:50 10 of (c) 90
490
Hormone panel 1 1:5 20 80
Note: Sample dilution factor for culture media samples must be optimized
by the end user.