M65® ELISA (PEVIVA®)
Apoptosis + Necrosis.
Human intermediate filament protein cytokeratin 18 (K18).
Released from human epithelial cells.
Store samples at 2 – 8 °C up to 4 hours. For longer
periods, store samples frozen at -20 °or lower.
Samples can be freeze-thawed without loss of activity it is recommended that repeated freeze thawing should be avoided.
Median level 264 U/L with a range between 136 – 480 U/L.
95th percentile was 413 U/L.
Total soluble K18 (uncleaved and cleaved Keratin 18 representing total cell death (necrosis and apoptosis) was measured in 200 apparently normal Swedish blood donors. Male and female values showed a similar distribution and no age dependency. Based on the distribution the cut-off value for elevated K18 has been set at > 450 U/L.
|Product name||M65® ELISA (PEVIVA®)|
|Range||125 - 2000 U/L (The Units measured by the M65® ELISA are defined against a synthetic standard. 1 U/L = 1.24 pM.)|
|Incubation time||2 hours 20 minutes|
|Sample volume||25 µl|
serum, plasma (EDTA, Heparin, Citrate) and cell (epithelial cell
Human , primates, bovine
The assay uses two monoclonal antibodies directed to epitopes in the
The Standard provided with the kit contains recombinant material which is not comparable to the fragment measured in the sample and is not adequate for spiking recovery tests. The recovery for the recombinant K18 protein fragment observed in human serum/plasma will be higher than expected.
Recovery for human sera when diluted 1:2 in M65 Standard A (0 Units/L): 126 % (average) and 116 – 139 % (range).
No high dose “hook effect” occurs before 70 000 U/L.
Quantitative measurement of total soluble cytokeratin 18 (K18) released from dead cells (necrotic and apoptotic). The cells or tissues should be of human epithelial origin (e.g. kidney, gut, colon, lung or liver) expressing K18.
The M65® ELISA can be combined with the M30-Apoptosense® ELISA (PEVIVA Prod. No. 10010) for determination of cell death mode (apoptosis versus necrosis). Death mode can be determined in vitro and in serum from cancer patients (Kramer et al., Cancer Res. 2004).