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- Antibody Data
- Antigen structure
- References [1]
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- Validations
- Western blot [3]
- Immunohistochemistry [1]
- Other assay [2]
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- Product number
- MA5-16368 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Glypican 3 Monoclonal Antibody (SP86)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- Heat-mediated antigen retrieval is recommended prior to staining, using a 10mM citrate buffer, pH 6.0, for 10 minutes followed by cooling at room temperature for 20 min. Following antigen retrieval, incubate samples with primary antibody for 30 min at room temperature. A suggested positive control is placenta or liver carcinoma.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- SP86
- Vial size
- 500 µL
- Concentration
- 0.466 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
Submitted references Impaired Autophagy Response in Hepatocellular Carcinomas Enriches Glypican-3 in Exosomes, Not in the Microvesicles.
Koksal AR, Thevenot P, Aydin Y, Nunez K, Sandow T, Widmer K, Nayak L, Scott J, Delk M, Moehlen MW, Cohen AJ, Dash S
Journal of hepatocellular carcinoma 2022;9:959-972
Journal of hepatocellular carcinoma 2022;9:959-972
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of HepG2 Cell Lysate using anti-Glypican-3 Monoclonal Antibody (Product # MA5-16368). The recommened dilution for this antibody in western blot applications is 1:25.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of HepG2 Cell Lysate using anti-Glypican-3 Monoclonal Antibody (Product # MA5-16368). The recommened dilution for this antibody in western blot applications is 1:25.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using anti-Glypican 3 Monoclonal Antibody (SP86) (Product # MA5-16368) and a 66 kDa band corresponding to GPC3 was observed in Hep G2 and not in HeLa, MCF7 and T-47D which are reported to be negative. Membrane enriched extracts (30 µg lysate) of Hep G2 (Lane 1), HeLa (Lane 2), MCF7 (Lane 3) and T-47D (Lane 4) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of Glypican-3 using anti-Glypican-3 Monoclonal Antibody (Product # MA5-16368) in Placenta Tissue. The recommened dilution for this antibody in immunohistochemistry applications is 1:100.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Show immune-magnetic affinity isolation detect GPC3 in HCC cell culture-derived exosomes, not in the microvesicles. ( A ) Detection of exosomes and microvesicles in the extracellular vesicles isolated from HCC cell culture. First, EVs were isolated from HCC cell culture using by total exosome isolation reagent. Second, exosomes and microvesicles were selected from total EVs using CD63 or Annexin A1 antibodies conjugated magnetic beads. Immuno-affinity capture exosomes or microvesicles were stained with Alexa Fluor 488 labeled GPC3. Approximately 53.4% of CD63 conjugated magnetic beads show positive for GPC3 as compared to 0.9% of Annexin A1 conjugated beads. About 92.8% of CD63 beads show positive for CD81, as compared to 45.7% of Annexin A1 beads. ( B ) Western blot analysis shows the distribution of GPC3 in the total non-selected and selected EVs. ( C ) The bar graph represents the relative intensity of GPC3 proteins according to particle-specific protein bands in Western blot. The results are expressed as the mean intensity +- standard deviation and analyzed by Mann Whitney U -test. *P < 0.05, **P < 0.01.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Impaired autophagy promotes GPC3 accumulation in exosomes. CD63-GFP stable cell line was treated with increasing concentrations of Torin 1 or Bafilomycin A1 for 72 hours. ( A ) Demonstrate the expression of CD63-GFP in Torin 1 treated and Bafilomycin A1 treated by fluorescence microscopy. ( B ) Quantification of CD63-GFP expression in Torin 1 and Bafilomycin A1 treated culture by Image J software. ( C ) Fluorescence microscopy images show the expressions of GPC3, and Tsg 101 in Torin 1 and Bafilomycin A1 treated cells. ( D ) Western blot analysis shows autophagy induction by Torin 1 leads to GPC3 and CD63 degradation. Data indicate mean +- standard deviation and are analyzed by the Mann-Whitney U -test. P values were displayed as **P
