United Kingdom (UK)
Recombinant monoclonal antibody to Glypican 3. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma 15G1.
UniProt Accession Number of Target Protein: P51654
Alternative Name(s) of Target: GPC3; Glypican-3; GPC-3; GTR2-2; MXR7; OCI5; DGSX; Intestinal protein OCI-5
Immunogen: The original antibody was generated by immunizing BALB/c mice with a recombinant human GPC3 extracellular domain (ECD, amino acids of 1-547) fused to a C-terminal Flag (RADYKDDDDK).
Specificity: This antibody binds an epitope between amino acids 420 to 470 of the human glypican 3 (GPC3). This antibody cross reacts with cynomolgus monkey and rat GPC3. Glypican-3 is an oncofetal antigen that belongs to the glypican family of glycosyl-phosphatidylinositol-anchored heparin sulfate proteoglycans. It negatively regulates the hedgehog signaling pathway and positively regulates the canonical Wnt signaling pathway. Multiple studies have revealed that GPC-3 is overexpressed in hepatocellular carcinoma (HCC).
Application Notes: This antibody can bind human glypican 3 expressed on the surface of 293S cells, HepG2 X1 cells and JHH7 cells in a flow cytometric assay. This antibody reacts with human rat and cynomolgus monkey GPC3 in a flow cytometric assay. The binding characterization of this antibody to huGPC3, cynoGPC3, and HepG2 cells was done using ELISA and immunohistochemistry. (US9926377).
Antibody first published in: PMID:
Flow cytometry using anti-Glypican 3 antibody 15G1 (Ab03202). HepG2 cells were stained with the anti-unknown specificity antibody (Ab00178-23.0; isotype control, black line) or the rabbit IgG version of 15G1 (Ab03202-23.0, blue line) at a dilution of 1:100 for 1h at RT. After washing, the bound antibody was detected using a goat anti-rabbit IgG AlexaFluor® 488 antibody at a dilution of 1:1000, and the cells were analyzed using a FACSCanto flow-cytometer.
Western blot using anti-Glypican 3 antibody 15G1 (Ab03202). HepG2 cells lysate (35µg protein in RIPA buffer) was resolved on an SDS-PAGE gel, and blots were probed with the chimeric rabbit version of 15G1 (Ab03202-23.0) at 0.1µg/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1 hour was used, and protein was detected by chemiluminescence.
Immunofluorescence staining of HepG2 cells with anti-Glypican 3 antibody 15G1 (Ab03202). Immunofluorescence analysis of paraformaldehyde fixed HepG2 cells on Shi-fix™ coverslips stained with the chimeric rabbit IgG version of 15G1 (Ab03202-23.0) (1:100 dilution) for 1h followed by Alexa Fluor® 488 secondary antibody (1:1000 dilution), showing membrane staining. The nuclear stain is DAPI (blue). Panels show, from left-right, top-bottom, Ab03202-23.0, DAPI, merged channels, and an isotype control. The isotype control was an unknown specificity antibody (Ab00178-23.0) followed by staining with Alexa Fluor® 488 secondary antibody.