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Synthetic peptide conjugated to KLH derived from within residues 350 to the C-terminus of Human Isocitrate dehydrogenase.
Our Abpromise guarantee covers the use of ab94571 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 47 kDa (predicted molecular weight: 47 kDa).|
|IHC-P||Use a concentration of 5 µg/ml.|
|ICC/IF||Use a concentration of 5 µg/ml.|
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: IDH1 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: HepG2 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab94571 observed at 46 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab94571 was shown to specifically recognize IDH1 in wild-type HAP1 cells along with additional cross reactive bands. No band was observed when IDH1 knockout samples were examined. Wild-type and IDH1 knockout samples were subjected to SDS-PAGE. Ab94571 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
IHC image of Isocitrate dehydrogenase staining in Human normal liver formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab94571, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.