Mouse Anti-VEGF Recombinant Antibody (clone A4.6.1) (CAT#: PABW-130)

Figure 1 In vivo potency and efficacy of anti-VEGF antibodies in inhibiting HM-7 (human colorectal) xenografts.

A, dose-responsive inhibition of tumor growth by A4.6.1, Y0317, B20-4.1, and G6-31 (5, 0.5, and 0.1 mg/kg, twice weekly) in HM-7 bearing mouse was shown by the tumor size (mm³) as a function of time (days). Error bars represent the S.E. (n = 10).

Liang, W. C., Wu, X., Peale, F. V., Lee, C. V., Meng, Y. G., Gutierrez, J., ... & Fuh, G. (2006). Cross-species vascular endothelial growth factor (VEGF)-blocking antibodies completely inhibit the growth of human tumor xenografts and measure the contribution of stromal VEGF. Journal of Biological Chemistry, 281(2), 951-961.

Figure 2 Examination of tumor sections.

B, medium magnification images (bar = 500 μm) of pancreatic HPAC (a, e, and i) and colonic HM-7 (b, f, and j) xenografts treated with control antibody (a and b), anti-human VEGF, Avastin (A4.6.1 for HM-7) (e and f), or anti-mouse/human VEGF, G6-31 (i and j). All images are centered 1 mm from the tumor/host margin. High magnification images (bar = 50 μm) of HPAC (c, g, and k) and HM-7 (d, h, and l) tumor xenografts treated with control antibody (c and d), anti-human VEGF, Avastin (g and h), or anti-mouse/human VEGF, G6-31 (k and l). Compared with HM-7, HPAC xenografts are relatively stromarich (compare c and d). Control antibody-treated HPAC tumors have extensive stromal cells infiltration (arrowheads, c), with indistinct small-caliber vessels surrounded by connective tissue cells and matrix. Treatment of these tumors with anti-human (g) or anti-mouse/human VEGF (k) results in little change in tumor-stroma proportion. In contrast, control HM-7 tumors (d) have relatively sparse stroma, and large caliber thin walled vessels (arrow). Treatment with anti-human VEGF (h) reduces vessel diameter and increases the relative amount of stromal matrix and cells around remaining vessels. Similar but more dramatic changes occur when HM-7 tumors are treated with anti-mouse/human VEGF, G6-31 (l). Images c, d, g, h, and k are centered 1 mm from the tumor/host margin (left in the images); image l is centered 200 μm from the tumor/host margin (in this sample, only necrotic tumor is present at 1 mm from the host margin).

Liang, W. C., Wu, X., Peale, F. V., Lee, C. V., Meng, Y. G., Gutierrez, J., ... & Fuh, G. (2006). Cross-species vascular endothelial growth factor (VEGF)-blocking antibodies completely inhibit the growth of human tumor xenografts and measure the contribution of stromal VEGF. Journal of Biological Chemistry, 281(2), 951-961.

Figure 3 2C3 inhibits VEGF-induced phosphorylation of VEGFR2.

PAE/KDR cells were stimulated for 15 min with PBS, basic fibroblast growth factor (5 nm, 100 ng/ml), VEGF165 (5 nm, 210 ng/ml), A673-conditioned media (CM), or CM + the indicated IgG (100 nm, 15 μg/ml). The cells were than incubated in lysis buffer, and the receptor was immunoprecipitated, separated by SDS-PAGE under reducing conditions, transferred to PVDF membranes, probed with mouse antiphosphotyrosine antibody (4G10), and developed under standard chemiluminescence conditions. The membranes were then stripped and reprobed with the immunoprecipitating IgG (T014) to determine the level of receptor protein in each lane.

Brekken, R. A., Overholser, J. P., Stastny, V. A., Waltenberger, J., Minna, J. D., & Thorpe, P. E. (2000). Selective inhibition of vascular endothelial growth factor (VEGF) receptor 2 (KDR/Flk-1) activity by a monoclonal anti-VEGF antibody blocks tumor growth in mice. Cancer research, 60(18), 5117-5124.

Figure 1 Mouse Anti-VEGF Recombinant Antibody (PABW-130) in WB

Western blot analysis was performed with 0.1 µg (lane 1), 0.3 µg(lane 2), and 0.6 µg (lane 3) recombinant human VEGF onto a 12% Tris-HCl polyacrylamide gel. Proteins were transferred to a CN membrane and blocked with 5% skim milk for at least 1 hour. Membranes were probed with PABW-130 at 2ng/μL overnight at 4°C on a rocking platform. Then probed with Goat anti-Mouse IgG HRP secondary antibody at a dilution of 1:3,000 for one hour. Chemiluminescent was detected.

Figure 2 Mouse Anti-VEGF Recombinant Antibody (PABW-130) in Dot Blot

Dot Blot analysis of PABW-130 was performed by coating with human VEGF antigen.
PABW-130 incubation concentration: 2 ng/μL.
The secondary antibody: HRP-Anti-Mouse IgG (H+L)

Figure 3 Mouse Anti-VEGF Recombinant Antibody (PABW-130) in ELISA

ELISA analysis of PABW-130 was performed by coating with recombinant human VEGF antigen. Then blocked with BSA, and incubated with PABW-130 at a starting concentration of 2.5 µg/mL. The HRP-conjugated Goat anti-Mouse IgG as a secondary antibody. Detection was performed using TMB substrate and stopped with sulfuric acid. The absorbances were read on a spectrophotometer at 450 nm.