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Our Abpromise guarantee covers the use of ab8934 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ELISA||1/8000 - 1/32000.|
|ICC/IF||Use a concentration of 1 µg/ml.|
|IHC-P||1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|WB||1/500 - 1/2000. Detects a band of approximately 52 kDa (predicted molecular weight: 52 kDa). We recommend overnight blocking with BSA solution at 4C, and incubating with the primary antibody overnight.|
|IHC-Fr||1/1000. PubMed: 17405874|
|IHC-FoFr||Use at an assay dependent concentration.|
Lane 1: 1/500
Lane 2: 1/1000
ab8934 staining PPARα in serum starved HepG2 cells (ab7900) treated with telmisartan (ab120831), by ICC/IF. Increase in PPARα expression correlates with increased concentration of telmisartan, as described in literature.
The cells were incubated at 37°C for 6h in media containing different concentrations of ab120831 (telmisartan) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab8934 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
ab8934 staining of PPAR alpha in rat brain (ab29475) sections, highlighting cytoplasmic staining in ependymal cells and neurons in frontal cortex. Top image shows subventricular zone (svz) of lateral ventrical (exit point of progenitor olfactory neurones); lower image shows frontal cortex in the same section. Cytoplasmic staining is also observed in the corpus callosum (top image) and in dendritic fields of the cortex. Formalin/PFA-fixed paraffin-embedded sections of rat brain tissue (ab29475) were incubated with ab8934 (1/200) for 1 hour. Antigen retrieval was performed by heat induction in citrate buffer pH 6.0. Please see accompanying abreview for additional information.
ICC/IF image of ab8934 stained HepG2 cells (ab7900). The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum (ab7481) / 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 (ab8934, 1µg/ml) overnight at +4°C. 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) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
ab8934 staining PPAR in Rat brain tissue (ab29475) sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with acetone, permeablized with methanol and blocked with 5% BSA for 1 hour at 37°C. The sample was incubated with primary antibody (1/100 in PBS) at 4°C for 18 hours. An Alexa Fluor® 488-conjugated Goat anti-rabbit polyclonal (1/200) (ab150077) was used as the secondary antibody.
ab8934 staining PPAR alpha in mouse liver tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent formaldehyde fixation before enzymatic antigen retrieval with Protease 0.05% in PBS for 5 min and then blocking with 5% serum was performed for 20 minutes at 20°C. The primary antibody was diluted 1/50 and incubated with sample in Tris + 5% normal goat serum for 1 hour at 20°C. A Biotin conjugated goat polyclonal to rabbit IgG was used at dilution at 1/500 as secondary antibody. Images show nuclear staining in hepatocytes (perfusion-fixed mouse, 10 and 40x microscope magnification).
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