Human Anti-FLT3 Recombinant Antibody (clone NC7) (CAT#: HPAB-0183CQ)

Figure 1 IMC-NC7 binds wild-type and mutant FLT3 and competes with FLT3 ligand for FLT3 binding.

IMC-NC7 binding to FLT3 was measured by ELISA, using plates coated with FLT3-Fc fusion protein and detected with horseradish peroxidase–conjugated anti-human κ antibody.

Piloto, O., Levis, M., Huso, D., Li, Y., Li, H., Wang, M. N.,... & Zhu, Z. (2005). Inhibitory anti-FLT3 antibodies are capable of mediating antibody-dependent cell-mediated cytotoxicity and reducing engraftment of acute myelogenous leukemia blasts in nonobese diabetic/severe combined immunodeficient mice. Cancer Research, 65(4), 1514-1522.

Figure 2 IMC-NC7 binds wild-type and mutant FLT3 and competes with FLT3 ligand for FLT3 binding.

IMC-NC7 binding to cell surface wild-type FLT3 on EOL-1 cells and mutant FLT3 on Ba/F3-ITD cells was determined by fluorescence-activated cell-sorting analysis using a phycoerythrin-conjugated anti-human secondary antibody.

Piloto, O., Levis, M., Huso, D., Li, Y., Li, H., Wang, M. N.,... & Zhu, Z. (2005). Inhibitory anti-FLT3 antibodies are capable of mediating antibody-dependent cell-mediated cytotoxicity and reducing engraftment of acute myelogenous leukemia blasts in nonobese diabetic/severe combined immunodeficient mice. Cancer Research, 65(4), 1514-1522.

Figure 3 IMC-NC7 blocks FLT3 ligand–mediated activation of wild-type FLT3 and FLT3 ligand–independent activation of mutant FLT3 along with downstream signaling.

Serum-starved (A) Ba/F3-FLT3 cells, (B) 32D/FLT3-ITD, and (C) Ba/F3-FLT3(D835γ) cells (10 × 106 cells/mL) were incubated with PBS, control IgG, or IMC-NC7 for 1 to 2 hours. Ba/F3-FLT3 cells were stimulated with 50 ng/mL FLT3 ligand for 15 minutes. Immunoprecipitates and total protein extracts were resolved by 8% or 10% SDS-PAGE, respectively, and subjected to immunoblot analysis with the indicated phosphospecific antibodies. The same blots were then stripped and reprobed with protein-specific antibodies.

Piloto, O., Levis, M., Huso, D., Li, Y., Li, H., Wang, M. N.,... & Zhu, Z. (2005). Inhibitory anti-FLT3 antibodies are capable of mediating antibody-dependent cell-mediated cytotoxicity and reducing engraftment of acute myelogenous leukemia blasts in nonobese diabetic/severe combined immunodeficient mice. Cancer Research, 65(4), 1514-1522.

Figure 4 IMC-NC7 inhibits FLT3 ligand–mediated wild-type FLT3 activation better than IMC-EB10.

EM3 cells (10 × 106 cells/mL) were incubated with IMC-EB10 or IMC-NC7 for 45 minutes and then treated with FLT3 ligand for 15 minutes. Immunoprecipitates were resolved by 8% SDS-PAGE and subjected to immunoblot analysis with phosphospecific antibody. The same blot was then stripped and reprobed with FLT3-specific antibody.

Piloto, O., Levis, M., Huso, D., Li, Y., Li, H., Wang, M. N.,... & Zhu, Z. (2005). Inhibitory anti-FLT3 antibodies are capable of mediating antibody-dependent cell-mediated cytotoxicity and reducing engraftment of acute myelogenous leukemia blasts in nonobese diabetic/severe combined immunodeficient mice. Cancer Research, 65(4), 1514-1522.

Figure 5 IMC-EB10 and IMC-NC7 mediate antibody-dependent cell-mediated cytotoxicity against Ba/F3-FLT3 and Ba/F3-ITD cells.

Ba/F3-FLT3 (A) and Ba/F3-ITD (B) cells were labeled with 51Cr and incubated with control IgG, IMC-C225, IMC-EB10, or IMC-NC7 along with an increasing titration of NK cells for 6 hours in triplicates. Released 51Cr was then measured by a gamma counter and percent lysis determined. SDS was used to determine 100% cell lysis.

Piloto, O., Levis, M., Huso, D., Li, Y., Li, H., Wang, M. N.,... & Zhu, Z. (2005). Inhibitory anti-FLT3 antibodies are capable of mediating antibody-dependent cell-mediated cytotoxicity and reducing engraftment of acute myelogenous leukemia blasts in nonobese diabetic/severe combined immunodeficient mice. Cancer Research, 65(4), 1514-1522.

Figure 1 FLT3 Antibody (HPAB-0183CQ) in WB

Western blot analysis was performed by loading 1 µg (lane 1) and 1 µg (lane 2) recombinant human FLT3 antigen 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 TLT3 antibody (HPAB-0183CQ) at a dilution of 1:1000 overnight at 4°C on a rocking platform. Then probed with Goat anti-Human IgG HRP secondary antibody at a dilution of 1:8,000 for one hour. Chemiluminescent was detected.

Figure 2 FLT3 Antibody (HPAB-0183CQ) in ELISA

ELISA analysis of HPAB-0183CQ was performed by coating with recombinant human FLT3 antigen. Then blocked with BSA, and incubated with TLT3 antibody (HPAB-0183CQ) at a starting concentration of 1µg/mL. The HRP-conjugated goat anti-human 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.

Figure 1 Anti-FLT3 Antibody (HPAB-0183CQ) in ELISA

ELISA analysis of HPAB-0183CQ was performed by coating with FLT3 Protein, Human, Recombinant (His Tag).

The secondary antibody: HRP-Anti-Human IgG (H+L)