MA5-16368

antibody from Invitrogen Antibodies

Targeting: GPC3 DGSX, OCI-5, SDYS, SGB, SGBS, SGBS1

Western blot

Antibody data

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Validation data

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

GPC3 protein structure - MA5-16368 shown in red.

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.

Supportive validation

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