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Baculovirus-expressed recombinant fragment: MADYSAALDP AYTTLEFENV QVLTMGNDTS PSEGTNLNAP NSLGVSAL, corresponding to amino acids 3-49 of Human HNF4
Our Abpromise guarantee covers the use of ab41898 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 53 kDa (predicted molecular weight: 53 kDa).|
|ELISA||Use a concentration of 0.1 µg/ml.|
|IP||Use at an assay dependent dilution.|
|IHC-P||Use a concentration of 10 - 20 µg/ml.|
|ICC/IF||Use a concentration of 1 µg/ml.|
|Flow Cyt||Use 1-2µg for 106 cells.|
|ChIP||Use at an assay dependent dilution. PubMed: 20463007|
|Gel supershift assays||Use at an assay dependent dilution.|
ICC/IF image of ab41898 stained HepG2 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 (ab41898, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) (ab150113) 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.
Overlay histogram showing HepG2 cells stained with ab41898 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum (ab7481) / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab41898, 2µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a significantly decreased signal in HepG2 cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Tween used under the same conditions.
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