Evaluation of an enzyme-linked immunosorbent assay to actin in cultured fibroblasts.
The amount of actin in cultured human skin fibroblasts was determined by an enzyme-linked immunosorbent assay. Absorbance values (OD), resulting from specific binding of an antiactin monoclonal antibody to intracellular actin, were converted to cell concentrations (pg/cell) from an actin standard curve. Actin concentration for cultured human skin fibroblasts by this technique was estimated to be 55.5 picograms/cell and constituted approximately 3.9% of the total cellular protein. This assay procedure offers the following advantages: (1) It is time efficient, can be completed in 2-3 hours; (2) cells are unaltered except for membrane permeabilization; (3) the sensitivity is equal to or greater than previous procedures involving gel electrophoresis; and (4) the assay is easy and inexpensive to perform.
Vande Berg
J S
JS
San Diego Veterans Administration Medical Center, CORE Electron Microscopy Unit, San Diego, Ca. 92161.
Disharoon
D R
DR
Poolman
W L
WL
Rudolph
R
R
D000199:Actins / Q000032:analysis* / Q000592:standards; D002452:Cell Count; D002469:Cell Separation; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D005069:Evaluation Studies as Topic; D005347:Fibroblasts / Q000032:analysis*; D006801:Humans; D012015:Reference Standards
D000199:Actins