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Full length native protein (purified) corresponding to Cow SDHA. Purified mitochondrial complex II (Cow).
Our Abpromise guarantee covers the use of ab14715 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC||Use a concentration of 0.2 µg/ml. Requires heat-induced antigen retrieval where aldehydes are used as fixatives. Use 20min incubation at 90-100°C in 0.1 M Tris/HCl pH 9.5 with 5% urea (wt/vol).|
|IHC-Fr||Use at an assay dependent concentration.|
|Flow Cyt||Use a concentration of 1 µg/ml.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|WB||Use a concentration of 0.1 µg/ml. Detects a band of approximately 70 kDa (predicted molecular weight: 70 kDa).|
|IHC-P||Use at an assay dependent concentration. PubMed: 20484225|
Mitochondrial localization of complex II visualized by immunocytochemistry using ab14715. Cultured human embryonic lung-derived fibroblasts (strain MRC5) were fixed, permeabilized and then labeled with ab14715 (0.2 µg/ml) followed by an AlexaFluor® 488-conjugated-goat-anti-mouse IgG2a isotype specific secondary antibody (2 µg/ml).
ICC/IF image of ab14715 stained human HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab14715, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in Hek293, HepG2 and MCF7 cells.
Lane 1: Prestained Protein Ladder
Lanes 2-3: Fly mitochondrial fraction
Lane 4: Fly cytoplasmic fraction
Lane 5: Robin mitochondrial fraction
Lane 6: Robin cytoplasmic fraction
Lane 7: Mouse (AML12) mitochondrial fraction
Lane 8: Mouse (AML12) cytoplasmic fraction
Lane 9: Prestained Protein Ladder
Mitochondrial and cytoplasmic fractions were obtained from whole flies with Abcam Mitochondrial Isolation Kit for Tissue (ab110168) and from robin fibroblast and mouse hepatocyte cell lines with Abcam Mitochondrial Isolation Kit for Cells (ab110170).
ab14715 was used to stain SDHA at 0.1 μg/ml
HRP-conjugated Horse Anti-mouse IgG at 1 μg/ml was used as a secondary antibody.