Mouse Anti-DLL4 Recombinant Antibody (clone MMGZ01) (CAT#: HPAB-0211-WJ)

Figure 1 Generation and characterization of MMGZ0.

(A) The serum titer of the mouse after the impact immunization was measured by indirect ELISA. The mouse with serum titer reaching 1:200,000 was selected.

Xu, Z., Wang, Z., Jia, X., Wang, L., Chen, Z., Wang, S., ... & Wu, M. (2016). MMGZ01, an anti-DLL4 monoclonal antibody, promotes nonfunctional vessels and inhibits breast tumor growth. Cancer letters, 372(1), 118-127.

Figure 2 Generation and characterization of MMGZ0.

(C) Western blot assay of his-tagged rhDLL4 or his-tagged MIC-A detected by MMGZ01. MMGZ01 specifically detected the rhDLL4 but not the unrelated recombinant protein MIC-A, which was detected by rabbit anti-his antibody.

Xu, Z., Wang, Z., Jia, X., Wang, L., Chen, Z., Wang, S., ... & Wu, M. (2016). MMGZ01, an anti-DLL4 monoclonal antibody, promotes nonfunctional vessels and inhibits breast tumor growth. Cancer letters, 372(1), 118-127.

Figure 3 Specificity analysis of MMGZ01.

(A, B) Affinity of MMGZ01 for rhDLL4 was determined by BIAcore. Results for the binding of MMGZ01 to immobilized rhDLL4 showed that the association rate increased with increasing concentration of the MMGZ01. The detailed concentrations of MMGZ01 (from bottom to up) are 0.00125 nM, 0.005 nM, 0.02 nM, 0.078 nM, 0.3125 nM, 1.25 nM, 5 nM and 20 nM.

Xu, Z., Wang, Z., Jia, X., Wang, L., Chen, Z., Wang, S., ... & Wu, M. (2016). MMGZ01, an anti-DLL4 monoclonal antibody, promotes nonfunctional vessels and inhibits breast tumor growth. Cancer letters, 372(1), 118-127.

Figure 4 Specificity analysis of MMGZ01.

(C) Flow cytometry analysis of the binding of rhDLL4-expressing cells with MMGZ01. HEK293 cells transfected with rhDLL4 (ECD) or a control vector (control) were analyzed using a FACSCalibur flow cytometer. The cells were incubated with a mouse anti-rhDLL4 antibody (positive control) or MMGZ01 and an APC-labeled secondary antibody.

Xu, Z., Wang, Z., Jia, X., Wang, L., Chen, Z., Wang, S., ... & Wu, M. (2016). MMGZ01, an anti-DLL4 monoclonal antibody, promotes nonfunctional vessels and inhibits breast tumor growth. Cancer letters, 372(1), 118-127.

Figure 5 Specificity analysis of MMGZ01.

(D) Characterization of MMGZ01 by immunofluorescent staining of transfected cells. HEK293 cells transfected with rhDLL4 (ECD) were subsequently fixed and stained immunofluorescently with or without MMGZ01. Green: GFP; Red: APC.

Xu, Z., Wang, Z., Jia, X., Wang, L., Chen, Z., Wang, S., ... & Wu, M. (2016). MMGZ01, an anti-DLL4 monoclonal antibody, promotes nonfunctional vessels and inhibits breast tumor growth. Cancer letters, 372(1), 118-127.

Figure 6 Specificity analysis of MMGZ01.

(F) MMGZ01 blocks DLL4-mediated inhibition of HUVEC proliferation. HUVECs growing on a 96-well plate pre-coated with 1 μg/mL of rhDLL4 protein were treated with different concentration of MMGZ01 or vehicle control for 96 hours.

Xu, Z., Wang, Z., Jia, X., Wang, L., Chen, Z., Wang, S., ... & Wu, M. (2016). MMGZ01, an anti-DLL4 monoclonal antibody, promotes nonfunctional vessels and inhibits breast tumor growth. Cancer letters, 372(1), 118-127.

Figure 7 MMGZ01 combined with docetaxel inhibits cell proliferation, induces apoptosis and promotes non-productive angiogenesis in MDA-MB-231 tumors.

(A) IHC staining of Ki-67 (anti-Ki 67 antibody) for proliferation in paraffin sections of xenografted tumor.

Xu, Z., Wang, Z., Jia, X., Wang, L., Chen, Z., Wang, S., ... & Wu, M. (2016). MMGZ01, an anti-DLL4 monoclonal antibody, promotes nonfunctional vessels and inhibits breast tumor growth. Cancer letters, 372(1), 118-127.