Measurement method | Analytical technique | Evaluated parameter | Pre-analytical factor | Threshold | Organ | References |
---|---|---|---|---|---|---|
Spectrophotometry | DC protein assay | Concentration determined based on standard curve | N/A | N/A | Colon, kidney | |
BCA protein assay | ||||||
Electrophoresis | Western blot | PCNA detection | Fixation | – | Colon | [57] |
Comparative evaluation of reactivity of fresh and FFPE using antibodies against GAPDH, tropomyosin, vinculin and myosin | – | – | Sheep tissue from skeletal muscle, liver, human hyperplastic thyroid tissue | [59] | ||
SDS-PAGE and silver staining | Size distribution | Sample age | High quality proteins are feasible to extract from 14Â years samples | Liver | [60] | |
N-cadherin and phospo-ERK detection | ||||||
2D-PAGE | Comparison of 2D-PAGE gel protein profiles | Time to freeze | 30Â min | Kidney | [61] | |
Immunoblotting | P-p27 detection | – | – | Cell culture | [62] | |
Mass spectrometry | LC–MS/MS analysis | Comparative analysis of peptide hits between fresh-frozen and FFPE samples | Fixation | – | Muscle | [59] |
Protein overlap between fresh and FFPE tissue sections | Fixation | – | Kidney | [58] | ||
Capillary isoelectric focusing coupled with RP LC–MS/MS | – | Storage time | From 7 years fewer distinct peptides and proteins were identified but the normalised expression values of actin, desmin and progesterone receptor were consistent until 12 years | Mesenchyme | [63] | |
Protein microarray | RPPAs | Evaluation of increase and decrease percentage of phosphoproteins | Time to fixation | 20Â min | Uterus, colon, lung, ovary, breast, lymph node | [64] |