rabbit polyclonal anti human anti gpc3  (Proteintech)

Journal: Molecular Medicine Reports

Article Title: Enhanced specific antitumor immunity of dendritic cells transduced with the glypican 3 gene and co-cultured with cytokine-induced killer cells against hepatocellular carcinoma cells

doi: 10.3892/mmr.2015.3239

Figure Lengend Snippet: Transduction efficiency in transduced DCs. (A) DCs-GPC3 observed using an inverted fluorescence microscope (magnification, ×100). (B) Transduction efficiency in DCs. Green fluorescent protein expression was evaluated using flow cytometry 24 h after gene transduction. (C) Reverse transcription-polymerase chain reaction analysis confirmed the mRNA expression of GPC3, using DCs-pcDNA3 and DCs as the controls. (D) Western blot analysis confirmed the protein expression of GPC3. Lanes: 1, DCs-pcDNA3; 2, DCs; 3: DCs-GPC3. DCs, dendritic cells; GPC3, glypican 3.

Article Snippet: A primary rabbit anti-human polyclonal antibody to GPC3 (sc-11395; 1:500) or an endogenous control β-actin (sc-7210; 1:500) were incubated with the membranes first and then with horseradish peroxidase-conjugated goat anti-rabbit IgGFc secondary antibodies (sc-2004; 1:2,000) (All antibodies from Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA).

Techniques: Transduction, Fluorescence, Microscopy, Expressing, Flow Cytometry, Reverse Transcription, Polymerase Chain Reaction, Western Blot

Journal: Molecular Medicine Reports

Article Title: Enhanced specific antitumor immunity of dendritic cells transduced with the glypican 3 gene and co-cultured with cytokine-induced killer cells against hepatocellular carcinoma cells

doi: 10.3892/mmr.2015.3239

Figure Lengend Snippet: Phenotypic analysis of cultured mature DCs measured by flow cytometry. Cell surface markers were detected in mature DCs, DCs transduced with either empty vector or GPC3. (A) Expression levels of CD80, CD83, CD86 and HLA-DR, detected by flow cytometry. (B) No statistically significant differences were observed in the surface phenotypic expression of mature DCs, DCs-pcDNA3 and DCs-GPC3. Data are expressed as the mean ± standard error of the mean from three independent experiments. DCs, dendritic cells; GPC3, glypican 3; CD, cluster of differentiation; HLA, human leukocyte antigen.

Article Snippet: A primary rabbit anti-human polyclonal antibody to GPC3 (sc-11395; 1:500) or an endogenous control β-actin (sc-7210; 1:500) were incubated with the membranes first and then with horseradish peroxidase-conjugated goat anti-rabbit IgGFc secondary antibodies (sc-2004; 1:2,000) (All antibodies from Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA).

Techniques: Cell Culture, Flow Cytometry, Transduction, Plasmid Preparation, Expressing

Journal: Molecular Medicine Reports

Article Title: Enhanced specific antitumor immunity of dendritic cells transduced with the glypican 3 gene and co-cultured with cytokine-induced killer cells against hepatocellular carcinoma cells

doi: 10.3892/mmr.2015.3239

Figure Lengend Snippet: Flow cytometric phenotypic analysis of CIKs 14 days after culturing. Treatment with CIKs co-cultured with DCs or DCs-pcDNA3 significantly increased the rate of CD3 + CD8 + and CD3 + CD56 + compared with the autologous CIKs alone ( * P<0.01, vs. CIKs). Furthermore, treatment with CIKs co-cultured with DCs-GPC3 significantly increased the rate of CD3 + CD8 + and CD3 + CD56 + compared with DCs-CIKs and DCs-pcDNA3-CIKs ( # P<0.05, vs. DCs-GPC3-CIKs). Data are expressed as the mean ± standard error of the mean from three independent experiments. DCs, dendritic cells; CD, cluster of differentiation; CIKs, cytokine-induced killer cells.

Article Snippet: A primary rabbit anti-human polyclonal antibody to GPC3 (sc-11395; 1:500) or an endogenous control β-actin (sc-7210; 1:500) were incubated with the membranes first and then with horseradish peroxidase-conjugated goat anti-rabbit IgGFc secondary antibodies (sc-2004; 1:2,000) (All antibodies from Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA).

Techniques: Cell Culture

Journal: Molecular Medicine Reports

Article Title: Enhanced specific antitumor immunity of dendritic cells transduced with the glypican 3 gene and co-cultured with cytokine-induced killer cells against hepatocellular carcinoma cells

doi: 10.3892/mmr.2015.3239

Figure Lengend Snippet: Intracellular IFN-γ-secreting CIKs determined by flow cytometry. IFN-γ secreted by DCs-pcDNA3-CIKs and DCs-CIKs was significantly higher compared with the CIKs alone ( * P<0.01, vs. CIKs). IFN-γ secreted by DCs-GPC3-CIKs was significantly higher compared with the DCs-pcDNA3-CIKs and DCs-CIKs ( # P<0.05, vs. DCs-GPC3-CIKs). Data are expressed as the mean ± standard error of the mean from three independent experiments. DCs, dendritic cells; CIKs, cytokine-induced killer cells; GPC3, glypican 3; IFN, interferon.

Article Snippet: A primary rabbit anti-human polyclonal antibody to GPC3 (sc-11395; 1:500) or an endogenous control β-actin (sc-7210; 1:500) were incubated with the membranes first and then with horseradish peroxidase-conjugated goat anti-rabbit IgGFc secondary antibodies (sc-2004; 1:2,000) (All antibodies from Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA).

Techniques: Flow Cytometry

Journal: Molecular Medicine Reports

Article Title: Enhanced specific antitumor immunity of dendritic cells transduced with the glypican 3 gene and co-cultured with cytokine-induced killer cells against hepatocellular carcinoma cells

doi: 10.3892/mmr.2015.3239

Figure Lengend Snippet: Cytotoxicity measured by a lactate dehydrogenase assay, using GPC3-expressing HepG2 cells as targets. The specific CIKs responses against HepG2 cells were investigated at effector to target ratios of 20:1 and 50:1. The cell lysis percentage in DCs-pcDNA3-CIKs and DCs-CIKs was higher compared with CIKs alone ( * P<0.01, vs. CIKs). DCs-GPC3-CIKs presented significantly greater lysis capacity against HepG2 cells, when compared with DCs-pcDNA3-CIKs and DCs-CIKs ( # P<0.01, vs. DCs-GPC3-CIKs). Data are expressed as the mean ± standard error of the mean from three independent experiments. DCs, dendritic cells; CIKs, cytokine-induced killer cells; GPC3, glypican 3.

Article Snippet: A primary rabbit anti-human polyclonal antibody to GPC3 (sc-11395; 1:500) or an endogenous control β-actin (sc-7210; 1:500) were incubated with the membranes first and then with horseradish peroxidase-conjugated goat anti-rabbit IgGFc secondary antibodies (sc-2004; 1:2,000) (All antibodies from Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA).

Techniques: Lactate Dehydrogenase Assay, Expressing, Lysis

Journal: Molecular Medicine Reports

Article Title: Enhanced specific antitumor immunity of dendritic cells transduced with the glypican 3 gene and co-cultured with cytokine-induced killer cells against hepatocellular carcinoma cells

doi: 10.3892/mmr.2015.3239

Figure Lengend Snippet: Inhibitory effects of effector cells was observed on HepG2 cell-induced tumor growth in tumor-bearing nude mice. (A) Tumor volume and (B) tumor weight in the DCs-GPC3-CIKs were found to be the smallest, followed by DCs-CIKs, CIKs and PBS. Data are expressed as the mean ± standard error of the mean from three independent experiments. * P<0.05 and ** P<0.01, vs. PBS group. DCs, dendritic cells; CIKs, cytokine-induced killer cells; GPC3, glypican 3; PBS, phosphate-buffered saline.

Article Snippet: A primary rabbit anti-human polyclonal antibody to GPC3 (sc-11395; 1:500) or an endogenous control β-actin (sc-7210; 1:500) were incubated with the membranes first and then with horseradish peroxidase-conjugated goat anti-rabbit IgGFc secondary antibodies (sc-2004; 1:2,000) (All antibodies from Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA).

Techniques: Saline