Table 1.

Preparation and analysis of secreted proteins

Mediuma	Incubation time (hours)	Conf.b (%)	Protein Recoveryc	Protein concentrationd	Analyzing method	Cell type	References
Argine and lysine-free	24	80	Filtration	MWCO filter	SILAC-SDS-PAGE-LC-MS/MS	Pancreatic duct epithelial ceUs (HPDE)	Gronborg et al. [19]
Serum-free						Pancreatic ductal adenocarcinoma cells (Panl)
Methionine and cysteine-free	2	NA	Filtration	Precipitation (ethanol)	LC-MS/MS	Hepathocarcinoma cells (HepG2)	Zwickl et al. [20]
Serum and phenol-free	24–18	NA	Centrifugation	Ultrafiltration	2DE-MALDI/MS	Murine myeloid cells (J774)	Chevallet et al. [13]
				Adsorption	2DE-LC-MS/MS
				Phase extraction (phenol)
				Precipitation (TCA)
Serum and phenol-free	72	75	Centrifugation	MWCO filter	ICAT-LC-MS/MS	Prostate cancer cells (LNCaP)	Martin et al. [80]
Serum-free	48	60–70	Centrifugation	MWCO filter	2DE-MALDI/MS	Colon carcinoma cells (SW480)	Volmer et al. [12]
			Filtration
Serum-free	48	NA	Centrifugation	Precipitation (TCA)	2DE-MALDI/MS	Glioblastoma cells (LN-Z308)	Khwaja et al. [18]
					ICAT-2DE-LC-MS/MS
Serum-free	36	60-70	Centrifugation	MWCO filter	SDS-PAGE-LC-MS/MS	Melanoma cells (UM)	Pardo et al. [16]
			Filtration	Precipitation (TCA/acetone)
Serum-free	30	60-70	Filtration	SPE (tC2)	LC-MS/MS	Melanoma cells (WM 266-4)	Mbeunkui et al. [14]
				Speed vacuum		Osteosarcoma cells (OHS)
						Breast cancer cells (MA11)
Serum-free	16	60	Centrifugation	SPE (tC2)	LC-MS/MS	Breast cancer cells (MCF10A, MCF10AT, MCF10 DCIS.com, MCF10CA cl. D)	Mbeunkui et al. [15]
			Filtration	Speed vacuum	SDS-PAGE-Immunoassay	Human tumor xenografts (MT-1)
				MWCO filter

^aModification of cell growth medium. Components added or removed from the medium

^bCell confluence in the growth medium prior to harvesting

^cMethods for recovering secreted proteins free of floating or dead cells

^dMethods for concentrating proteins from the growth medium