Fatty Acid Oxidation Human Flow Cytometry Kit (ab118183) contains antibodies against key enzymes of the mitochondrial fatty acid oxidation pathway for flow cytometry. The assay combines the power of single cell analysis obtained from flow cytometry with the specificity of antibody-based immunostaining to quantify protein levels in cultured cells. Cells are harvested and fixed/permeabilized in suspension, targets of interest are detected indirectly with highly specific well-characterized monoclonal antibodies that are then labeled with fluorescent antibodies.
Spin down the contents of the antibody vials upon receipt of the kit. Store all components upright at 4°C. 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 Flow Cytometry Kit (ab118183)
Antibody specificity demonstrated by immunocytochemistry. Cells were processes with the Flow cytometry protocol as explained above. 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.
Antibody specificity demonstrated by flow cytometry. Control (blue) and deficient cell lines (red) were targeted with the FAO antibody panel against ACADVL, ACADM and HADHA. Background fluorescence was determined with the supplied negative control (black). After background subtraction, the ACADVL deficient cell line shows a 93% reduction in the level of the ACADVL protein, the ACADM deficient line shows a 57% reduction in the level of the ACADM protein and the LCHAD deficient cell line shows a 50% reduction of in the level of the HADHA protein. The deficient cell lines characterized protein changes are: (1) ACADVL:p.[N122D], (2) ACADM:p[K304E and (3) HADHB:p[R61H];[R247H].
Cook KL et al. Endoplasmic Reticulum Stress Protein GRP78 Modulates Lipid Metabolism to Control Drug Sensitivity and Antitumor Immunity in Breast Cancer. Cancer Res76:5657-5670 (2016).
Read more (PubMed: 27698188) »