Glypican 3/GPC3 Antibody

• Catalog Number: orb692158
• Category: Antibodies
• Description: Glypican 3/GPC3 Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, FC, WB
• Reactivity: Human
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived human Glypican 3/GPC3 recombinant protein (Position: D32-A535).
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Dilution range: Western blot, 0.25-0.5μg/ml Flow Cytometry, 1-3μg/1x106 cells Direct ELISA, 0.1-0.5μg/ml
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 66 kDa
• UniProt ID: P51654
• Storage: Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
• Alternative names: Glypican-3; GTR2-2; Intestinal protein OCI-5; MXR7
• Note: For research use only
• Application notes: Tested Species: In-house tested species with positive results. Other applications have not been tested. Optimal dilutions should be determined by end users. Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
• Expiration Date: 12 months from date of receipt.

Western blot analysis of Glypican 3/GPC3 using anti-Glypican 3/GPC3 antibody (orb692158). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human HepG2 whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Glypican 3/GPC3 antigen affinity purified polyclonal antibody (Catalog # orb692158) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # orb90503) with Tanon 5200 system. A specific band was detected for Glypican 3/GPC3 at approximately 66 kDa. The expected band size for Glypican 3/GPC3 is at 66 kDa.

Flow Cytometry analysis of HepG2 cells using anti-Glypican 3/GPC3 antibody (orb692158). Overlay histogram showing HepG2 cells stained with orb692158 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Glypican 3/GPC3 Antibody (orb692158, 1 μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.