| Description | This product is a recombinant Mouse antibody provided by CreativeBiolabs. The antibody is specific for Glypican 3. It can be used for Glypican 3 detection in Enzyme-Linked Immunosorbent Assay. It was expressed in mammalian cells (293F or CHO) with antibody encoding genes and purified by affinity chromatography. Each lot of this antibody is quality control tested by SDS-PAGE and SEC-HPLC analysis. For highly sensitive assays, we recommend the ultrapurified form of the product, which has a lower endotoxin limit than standard antibody, less than 1 EU/mg or even 0.1 EU/mg. |
| Clonality | Monoclonal |
| Host Species | Mouse |
| Target Species | Human |
| Isotype | IgG |
| Expression Species | HEK293F or CHO Cell line |
| Conjugation | Unconjugated, also available for Biotin, HRP, FITC and PE-labeled form. |
| Purity | >95%, determined by SDS-PAGE and SEC-HPLC |
| Endotoxin | <1 EU/mg, determined by LAL method |
| Purification | Purified with Protein A or G affinity chromatography |
| Sterility | 0.2 μM filtered |
| Formulation | PBS, pH 7.4 |
| Preservation | No preservatives |
| Stabilizer | No stabilizers |
| Storage | Store at 4°C within one or two weeks. Store at -20°C for long term. Avoid repeated freeze/thaw cycles. Refer to the COA file for specifics. |
| Application | WB; ELISA |
| ELISA | Enzyme-Linked Immunosorbent Assay Protocol |
| WB | Western Blot Protocol |
| FC | Flow Cytometry Protocol |
Figure 1 Anti-GPC3 Monoclonal Antibody (V3S-1022-YC3236) in SDS-PAGE
SDS-PAGE analysis of V3S-1022-YC3236 in non-reduced (lane 1) and β-mercaptoethanol-reduced (lane 2) conditions. Gel stained for 30 minutes with Coomassie Blue. As a result of different β-mercaptoethanol-reduced proteins (Heavy chain and Light chain) migrate as about 50 kDa and 25 kDa, respectively. And non-reduced protein migrates as 130-180 kDa.
Figure 1 Anti-GPC3 Monoclonal Antibody (V3S-1022-YC3236) in SDS-PAGE
Figure 2 Anti-GPC3 Monoclonal Antibody (V3S-1022-YC3236) in WB
Western blot analysis of V3S-1022-YC3236 was performed by loading 2 µg onto a 12% Tris-HCl polyacrylamide gel in non-reduced (lane 1) and β-mercaptoethanol-reduced (lane 2) conditions.
Figure 2 Anti-GPC3 Monoclonal Antibody (V3S-1022-YC3236) in WB
Figure 3 Anti-GPC3 Monoclonal Antibody (V3S-1022-YC3236) in SEC-HPLC
The purity of V3S-1022-YC3236 was greater than 95% as determined by SEC-HPLC.
Column: 3 µm, 7.8 x 300 nm
Mobile phase: 150 mM Sodium Phosphate Buffer, pH 7.3
Detection: UV 280 nm
Injection: 25 µl
Figure 3 Anti-GPC3 Monoclonal Antibody (V3S-1022-YC3236) in SEC-HPLC
Figure 4 Anti-GPC3 Monoclonal Antibody (V3S-1022-YC3236) in WB
Western blot analysis was performed by loading 0.05 µg (lane 1) and 0/1 µg (lane 2) recombinant human GPC3 protein onto a 12% Tris-HCl polyacrylamide gel. Proteins were transferred to a CN membrane and blocked with 5% skim milk for at least 1 hour. Membranes were probed with GPC3 Therapeutic Antibody (V3S-1022-YC3236) at a dilution of 1:1000 overnight at 4°C on a rocking platform. Then probed with Goat anti-Human IgG HRP secondary antibody at a dilution of 1:8,000 for one hour. Chemiluminescent was detected.
Figure 4 Anti-GPC3 Monoclonal Antibody (V3S-1022-YC3236) in WB
Figure 5 Anti-GPC3 Monoclonal Antibody (V3S-1022-YC3236) in ELISA
ELISA analysis of V3S-1022-YC3236 was performed by coating with recombinant human GPC3 antigen. Then blocked with BSA, and incubated with GPC3 Therapeutic Antibody (V3S-1022-YC3236) at a starting concentration of 2.5 µg/mL. The HRP-conjugated goat anti-human IgG as a secondary antibody. Detection was performed using TMB substrate and stopped with sulfuric acid. The absorbances were read on a spectrophotometer at 450 nm.
Figure 5 Anti-GPC3 Monoclonal Antibody (V3S-1022-YC3236) in ELISA
