ab118182 uses quantitative immunocytochemistry to measure protein levels or post-translational modifications in cultured cells. Cells are fixed in a microplate and targets of interest are detected with highly specific, well-characterized monoclonal antibodies, and levels are quantified with IRDye®-labeled Secondary Antibodies. IR imaging and quantitation is performed using a LI-COR® Odyssey® or Aerius® system.
Plates are available in our ICE (In-Cell ELISA) Support Pack (ab111542) which can be bought seperately.
Upon receipt spin down the contents of the IRDye®-labeled Secondary Antibody tube and protect from light. Store all components upright at 4C. This kit is stable for at least 6 months from receipt.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
Immunocytochemistry - Fatty Acid Oxidation Human In-Cell ELISA Kit (ab118182)
Antibody specificity demonstrated by immunocytochemistry. Visualization under the microscope was carried out with a completely opened aperture and a very narrow field of visualization at 40x. Panel A shows control fibroblasts and panel B shows deficient fibroblasts. Left panel shows staining with anti-ACADVL ab, center panel with anti-ACADM ab and right panel with anti-HADHA ab. Note that although LCHAD#1 deficient had a characterized homozygous mutation, the antibody shows a mosaic pattern of staining with about 1/3 of the cells lacking HADHA staining and an overall 40% reduction of signal as observed by ICE.
Panel C shows levels of HADHA protein in two deficient cell lines: (1) HADHA:p.[E474Q] and (2) HADHB:p. [R61H];[R247H]. Note that although LCHAD#1 deficient has a partial deficiency as observed by ICC.