Anti-Human CLDN4 Recombinant Antibody (KM3900) (CAT#: TAB-501LC)

Figure 1 KM3900 binds to Claudin‐4 (CLDN4), but not to CLDN3, 5, 6, and 9. CLDN‐expressing CHO cells (CLDN3/CHO, CLDN4/CHO, CLDN5/CHO, CLDN6/CHO, and CLDN9/CHO) and vector‐transfected CHO (Vector/CHO) were generated.

The expression CLDNs were tagged myc/his sequence at the C‐terminus, so the expression was evaluated by anti‐myc or his antibodies. Control mouse IgG1 (gray histogram) and anti‐myc or anti‐his antibody (open histogram) reacted to CLDN4/CHO or Vector/CHO pretreated with 70% ethanol. The binding was detected by FITC‐conjugated antimouse antibody (a, left). Furthermore, CLDN4/CHO and Vector/CHO were subjected to fluorescence immunohistochemical analysis using anti‐myc antibody and the staining was visualized by confocal laser scanning microscope (white bar, 40 µm). The green staining represents the binding of anti‐myc antibody, which was detected with FITC‐conjugated antimouse antibody. The blue staining represents the cell nuclei staining with TOTO3 (a, right). Control mouse IgG2a (gray histogram) or KM3900 (open histogram) reacted to CLDN3, 4, 5, 6, 9, and Vector/CHO cells and the binding was detected by FITC‐conjugated antimouse antibody (b). The mRNA expressions of transfected genes were detected by RT‐PCR with sequence‐specific primers (b). Reverse transcriptase (–) was used as a negative control (RT–).

Suzuki, M., Kato‐Nakano, M., Kawamoto, S., Furuya, A., Abe, Y., Misaka, H.,... & Ando, H. (2009). Therapeutic antitumor efficacy of monoclonal antibody against Claudin‐4 for pancreatic and ovarian cancers. Cancer science, 100(9), 1623-1630.

Figure 2 Flow cytometry analysis of KM3900 against pancreatic cancer cells.

Cells were reacted with 5 µg/mL of KM3900 (open histogram) or control mouse IgG2a (gray histogram), and binding was detected by FITC‐conjugated antimouse antibody (a). Cell lysates derived from pancreatic cancer cells were separated with SDS‐PAGE and transferred to PVDF membrane. The bands show anti‐CLDN4 (3E2C1) or anti‐actin antibody reactions (b).

Suzuki, M., Kato‐Nakano, M., Kawamoto, S., Furuya, A., Abe, Y., Misaka, H.,... & Ando, H. (2009). Therapeutic antitumor efficacy of monoclonal antibody against Claudin‐4 for pancreatic and ovarian cancers. Cancer science, 100(9), 1623-1630.

Figure 3 Flow cytometry analysis of KM3900 against ovarian cancer cells.

Cells were reacted with 5 µg/mL of KM3900 (open histogram) or control mouse IgG2a (gray histogram), and binding was detected by FITC‐conjugated antimouse antibody (a). Cell lysates derived from ovarian cancer cells were separated with SDS‐PAGE and transferred to PVDF membrane. The bands show anti‐CLDN4 (3E2C1) or anti‐actin antibody reactions (b).

Suzuki, M., Kato‐Nakano, M., Kawamoto, S., Furuya, A., Abe, Y., Misaka, H.,... & Ando, H. (2009). Therapeutic antitumor efficacy of monoclonal antibody against Claudin‐4 for pancreatic and ovarian cancers. Cancer science, 100(9), 1623-1630.

Figure 1 Anti-CLDN4 Antibody (TAB-501LC) in SDS-PAGE

SDS-PAGE analysis of TAB-501LC in non-reduced (lane 1) and β-mercaptoethanol-reduced (lane 2) conditions. Gel stained for 30 minutes with Coomassie Blue. As a result of different β-mercaptoethanol-reduced proteins (Heavy chain and Light chain) migrate as about 50 kDa and 25 kDa, respectively. And non-reduced protein migrates as 130-180 kDa.

Figure 2 Anti-CLDN4 Antibody (TAB-501LC) in HPLC

The purity of TAB-501LC was greater than 95% as determined by SEC-HPLC.

Column: 3 µm, 7.8x300 nm
Mobile phase: 150 mM Sodium Phosphate Buffer, pH 7.3
Detection: UV280 nm
Injection: 15 µl