This antibody gave a positive signal in the following lysates: Human Kidney Tissue; Human Liver Tissue; HepG2 Whole Cell; HEK293 Whole Cell.
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pH: 7.40 Preservative: 0.02% Sodium azide Constituent: PBS Note: Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 µg/ml.
Use a concentration of 1 µg/ml. Predicted molecular weight: 66 kDa.
Cell surface proteoglycan that bears heparan sulfate. Inhibits the dipeptidyl peptidase activity of DPP4. May be involved in the suppression/modulation of growth in the predominantly mesodermal tissues and organs. May play a role in the modulation of IGF2 interactions with its receptor and thereby modulate its function. May regulate growth and tumor predisposition.
Highly expressed in lung, liver and kidney.
Involvement in disease
Defects in GPC3 are the cause of Simpson-Golabi-Behmel syndrome type 1 (SGBS1) [MIM:312870]; also known as Simpson dysmorphia syndrome (SDYS). SGBS is a condition characterized by pre- and postnatal overgrowth (gigantism) with visceral and skeletal anomalies.
Western blot - Anti-Glypican 3 antibody (ab101478)
Lane 1: Wild-type HAP1 whole cell lysate (20 µg) Lane 2: Glypican 3 knockout HAP1 whole cell lysate (20 µg) Lane 3: HepG2 whole cell lysate (20 µg)
Lanes 1 - 3: Merged signal (red and green). Green - ab101478 observed at 66 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab101478 was shown to specifically react with Glypican 3 in wild-type HAP1 cells as signal was lost in Glypican 3 knockout cells. Wild-type and Glypican 3 knockout samples were subjected to SDS-PAGE. ab101478 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1 μg/ml 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.
Western blot - Glypican 3 antibody (ab101478)
All lanes : Anti-Glypican 3 antibody (ab101478) at 1 µg/ml
Lane 1 : Human kidney tissue lysate - total protein (ab30203) Lane 2 : Human liver tissue lysate - total protein (ab29889) Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 66 kDa Observed band size: 66 kDa
ICC/IF image of ab101478 stained SKNSH 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 (ab101478, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit 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.