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Synthetic peptide conjugated to KLH derived from within residues 2450 to the C-terminus of Mouse Fatty Acid Synthase.
(Peptide available as ab25719.)
Our Abpromise guarantee covers the use of ab22759 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 1 µg/ml.|
|WB||Use at an assay dependent concentration. Detects a band of approximately 273 kDa (predicted molecular weight: 273 kDa).|
|IHC-P||Use at an assay dependent concentration.|
|IHC (PFA fixed)||Use a concentration of 2 µg/ml.|
|IHC-Fr||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration. PubMed: 21098489|
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Fatty Acid Synthase knockout HAP1 cell lysate (20 µg)
Lane 3: A549 cell lysate (20 µg)
Lane 4: Hu liver tissue lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab22759 observed at 250 kDa. Red - loading control, ab18058, observed at 124 kDa.
ab22759 was shown to specifically react with Fatty Acid Synthase in wild-type HAP1 cells. No band was observed when Fatty Acid Synthase knockout samples were examined. Wild-type and Fatty Acid Synthase knockout samples were subjected to SDS-PAGE. ab22759 and ab18058 (loading control to Vinculin) were diluted at 1 µg/ml and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
ab22759 staining Fatty Acid Synthase in Mouse colon tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1.5% serum for 30 minutes at 23°C; antigen retrieval was by heat mediation in Citra Plus solution. Samples were incubated with primary antibody (2μg/ml) for 14 hours at 4°C. A Biotin-conjugated Goat anti-rabbit IgG polyclonal (1/50) was used as the secondary antibody.
ICC/IF image of ab22759 stained human HEK 293 cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab22759, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions.
The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
Immunofluorescent staining for Fatty Acid Synthase in the rat striatum using Rabbit polyclonal to Fatty Acid Synthase (ab22759). Abundant staining was observed in the Striatum with lower levels of staining observed in the Corpus callosum. This is a montage of three pictures aquired using a X10 objective. ab22759 was used at 1/200 (2µg/ml) incubated overnight at room temperature.
Secondary antibody used was anti-rabbit Alexa Fluor® 488 at 1/1000 incubated for 2 hours at room temperature. Rat brain tissue was perfusion fixed with 4% PFA followed by overnight post-fixation in the same fixative, cryoprotected in 20% sucrose and frozen in OCT. 30µm coronal sections were cut on a cyrostat and immunohistochemistry performed by the 'free floating' technique.
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