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Synthetic peptide derived from C terminal of Human Glypican 3.
Our Abpromise guarantee covers the use of ab95363 in the following tested applications.
|ICC/IF||Use a concentration of 5 µg/ml.|
|Flow Cyt||Use at an assay dependent concentration.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|WB||Use at an assay dependent concentration. Predicted molecular weight: 66 kDa.|
|IHC-P||1/100. Perform heat mediated antigen retrieval (boil tissue section in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at RT for 20 min).|
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: GPC3 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HepG2 whole cell lysate (20 µg)
Lanes 1 - 3: Merged signal (red and green). Green - ab95363 observed at 70 kDa. Red - loading control, ab130007, observed at 125kDa.
ab95363 was shown to specifically react with Glypican 3 in wild-type HAP1 cells as signal was lost in GPC3 knockout cells. Wild-type and GPC3 knockout samples were subjected to SDS-PAGE. ab95363 and ab130007 (Mouse anti-vinculin loading control) were incubated overnight at 4°C at 1/1000 and 1/20000 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 1/10000 dilution for 1 hour at room temperature before imaging.
Immunohistochemical staining of human liver hepatocellular carcinoma with ab95363.
Flow cytometric analysis of rabbit anti-Glypican 3 (SP86) antibody ab98363 (1/100) in HEPG2 cellls (green) compared to negative control of rabbit IgG (blue).
ICC/IF image of ab95363 stained HepG2 cells. The cells were 4% formaldehyde 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 ab95363 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 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.
ab95363 has not yet been referenced specifically in any publications.