Apoptosis

Product name

M65® ELISA (PEVIVA®)

Apoptosis + Necrosis.
Human intermediate filament protein cytokeratin 18 (K18).
Released from human epithelial cells.

Cat-Nr.10020
Range125 - 2000 U/L (The Units measured by the M65® ELISA are defined against a synthetic standard. 1 U/L = 1.24 pM.)
Sensitivity11 U/L
Incubation time2 hours 20 minutes
Sample volume25 µl
Sample type

Human serum or plasma (EDTA, Citrate, Heparin plasma), containing
K18-reactive material released from K18 positive human cells.
Multiple freeze-thaw cycles of samples are well tolerated.
The same type of material i.e. serum or plasma collected by one method should be used for a specific project.
Mouse/Rat plasma can be used for measurement of human xenografts.
Cell culture supernatants from K18 positive (epithelial) cells or tissues.

Sample preparation

Fresh samples are stable for up to two days at 2 – 8 °C, for at least
9 months at -20 °C; and for at least two years when stored at -80 °C.

Reference values

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.

Species

Human, non-human primates, bovine.

Specificity

The assay uses two monoclonal antibodies directed to epitopes in the
284 – 396 region of the K18 protein. Soluble full length K18 as well as K18 fragments and protein complexes that expose these epitopes will be detected by the assay.

Tests96
MethodELISA
Wiederfindung

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.

Verduennung

Recovery for human sera when diluted 1:2 in M65 Standard A (0 Units/L): 126 % (average) and 116 – 139 % (range).
Recovery for human sera when diluted 1:1 to 1:10 in the M65 Sample Dilution Buffer: 106 % (average and 100 – 117 % (range).

Hook

No high dose “hook effect” occurs before 70 000 U/L.

Intended use

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).

Product informations

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