Development of an enzyme-linked immunosorbent assay (ELISA) for the specific detection of MEKK1 expression in cells.
BACKGROUND
MEKK1 is a 196-kDa serine-threonine kinase activated in response to a variety of stimuli, including EGF, lysophosphatidic acid, osmotic stress, UV light, and microtubule toxins. However, there are few reports about the expression level of MEKK1 in cancers.


METHODS
In this report, a direct competitive ELISA to quantify total MEKK1 in cell lines was developed.


RESULTS
The procedure showed a high sensitivity (detection limit: 0.17 ng/ml), good precision (coefficient of variation </=10.12) and acceptable linearity over a large range of MEKK1 concentrations (0.1-10,000 ng/ml). In a pilot study, this assay was used to quantify MEKK1 in different cell lines. In cancer cells, the range of MEKK1 is 0.02-85 ng/mg protein and its concentration was higher in pancreas cancer cells and umbilical vein cells than that in others.


CONCLUSIONS
This method can be used to measure the MEKK1 level of cell samples. The activity of MEKK1/JNK pathway in pancreas cancer and umbilical vein cell lines indicates that MEKK1 may be a potential target in interfering with pancreas cancer and angiogenesis.
Ding
Yan
Y
Department of Pharmacology, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, 1 Xian Nong Tan Street, Beijing 100050, China.
Huang
Dan
D
Chen
Xiao-Guang
XG
D000230:Adenocarcinoma / Q000201:enzymology / Q000473:pathology; D015153:Blotting, Western; D002460:Cell Line; D045744:Cell Line, Tumor; D002999:Clone Cells; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006367:HeLa Cells; D006801:Humans; D048728:MAP Kinase Kinase Kinase 1 / Q000032:analysis / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D008297:Male; D010190:Pancreatic Neoplasms / Q000201:enzymology / Q000473:pathology; D010865:Pilot Projects; D012680:Sensitivity and Specificity; D014471:Umbilical Veins / Q000166:cytology / Q000201:enzymology
D048728:MAP Kinase Kinase Kinase 1