Anti-Human GM2 (Monosialo-GM2) Recombinant Antibody (DMF10.167.4) (CAT#: TAB-170CT)

Figure 1 Analysis of DMF10.176.4 expression and effects on small intestine.

Intestine from untreated animals incubated ex vivo with control DMF10.167.4 and stained for bound hamster immunoglobulin.

Retter, M. W., Johnson, J. C., Peckham, D. W., Bannink, J. E., Bangur, C. S., Dresser, K., ... & Woda, B. (2005). Characterization of a proapoptotic antiganglioside GM2 monoclonal antibody and evaluation of its therapeutic effect on melanoma and small cell lung carcinoma xenografts. Cancer research, 65(14), 6425-6434.

Figure 2 GM2 expressed by RCC lines contributes to tumor-induced T-cell apoptosis.

Cell lines (2 × 10⁶ per tube) were stained with anti-GM2 antibody or IgG isotype control for 30 minutes at 4°C. T cells were then incubated with FITC-conjugated goat anti-hamster IgG antibody, and at least 20,000 events were acquired by FACScan. Percentage of GM2-positive cells in RCC cell lines SK-RC-54V, SK-RC-26B, and 0827 Lung Met (0827LM) and the normal epithelial cell line NKE. Similar results were observed in two more experiments.

Biswas, K., Richmond, A., Rayman, P., Biswas, S., Thornton, M., Sa, G., ... & Novick, A. (2006). GM2 expression in renal cell carcinoma: potential role in tumor-induced T-cell dysfunction. Cancer research, 66(13), 6816-6825.

Figure 3 GM2 expressed by RCC lines contributes to tumor-induced T-cell apoptosis.

Specificity of the hamster anti-human GM2 antibody (IgG) was determined by immunostaining of HPTLC plates spotted with a mixture of purified bovine brain-derived gangliosides: GM3, GM2, GM1, and GD3 (lane 1), purified GM2 (lane 2), and a mixture containing no GM2 (GM3, GM1, and GD3; lane 3). Plates were incubated with anti-GM2 antibody followed by counterstaining with secondary antibody (biotin-conjugated goat anti-hamster IgG) for 1 hour as described in Materials and Methods.

Biswas, K., Richmond, A., Rayman, P., Biswas, S., Thornton, M., Sa, G., ... & Novick, A. (2006). GM2 expression in renal cell carcinoma: potential role in tumor-induced T-cell dysfunction. Cancer research, 66(13), 6816-6825.

Figure 4 GM2 expressed by RCC lines contributes to tumor-induced T-cell apoptosis.

RCC cell lines SK-RC-54V (top) and SK-RC-26B (bottom) grown to confluence for 4 to 5 days were pretreated for 3 hours with either anti-GM2 antibody (GM2 Ab; 1 μg/mL) or control IgG antibody (IgG Ab; 1 μg/mL) before adding T cells. Following coculture for 72 hours, T cells were harvested for either TUNEL assay or trypan blue exclusion. The mean increase in T-cell death was significantly greater for lymphocytes cultured with tumor cells (with or without IgG control) versus medium. *, P < 0.03 versus T cells + medium. Addition of anti-GM2 antibody significantly reduced tumor-induced T-cell death. **, P < 0.02 versus T cells + SK-RC-26B (n = 3).

Biswas, K., Richmond, A., Rayman, P., Biswas, S., Thornton, M., Sa, G., ... & Novick, A. (2006). GM2 expression in renal cell carcinoma: potential role in tumor-induced T-cell dysfunction. Cancer research, 66(13), 6816-6825.

Figure 5 GM2 expression by a single-cell suspension from RCC digest.

Cells were stained for GM2 as already described. Cells were then observed under confocal microscope. GM2 is detected by the green fluorescence, and the nuclei of the cells, which are stained with DAPI, appear blue.

Biswas, K., Richmond, A., Rayman, P., Biswas, S., Thornton, M., Sa, G., ... & Novick, A. (2006). GM2 expression in renal cell carcinoma: potential role in tumor-induced T-cell dysfunction. Cancer research, 66(13), 6816-6825.