Mouse Anti-CD44 Recombinant Antibody (clone A3D8) (CAT#: HPAB-0141CQ)

Figure 1 H90 and A3D8 anti-CD44 mAbs inhibit proliferation of HL60, NB4, THP-1, and KG1a cell lines.

A total of 10⁵ cells/mL were seeded into 96-well plates and incubated in triplicate in the presence of specific anti-CD44 mAbs (2.5 μg/mL A3D8 or 20 μg/mL H90). Controls were cells incubated with isotypic control (20 μg/mL IgG1). At the indicated times, the viable cell number was determined using trypan blue dye exclusion

Charrad, R. S., Gadhoum, Z., Qi, J., Glachant, A., Allouche, M., Jasmin, C.,... & Smadja-Joffe, F. (2002). Effects of anti-CD44 monoclonal antibodies on differentiation and apoptosis of human myeloid leukemia cell lines. Blood, 99(1), 290-299.

Figure 2 Apoptotic cells, 3 days following addition of 2.5 μg/mL A3D8.

Fluorescent staining with acridine orange and ethidium bromide showed nuclei condensation and fragmentation that are apoptosis-specific. Cells at an early stage of apoptosis (without loss of membrane integrity) displayed a bright green–stained nucleus. Apoptic cells that have lost membrane integrity displayed bright orange–stained nuclei as a result of taking up ethidium bromide.

Charrad, R. S., Gadhoum, Z., Qi, J., Glachant, A., Allouche, M., Jasmin, C.,... & Smadja-Joffe, F. (2002). Effects of anti-CD44 monoclonal antibodies on differentiation and apoptosis of human myeloid leukemia cell lines. Blood, 99(1), 290-300.

Figure 3 Apoptotic cells, 3 days following addition of 2.5 μg/mL A3D8.

DNA strand breaks, which are apoptosis-specific, were labeled using TUNEL reaction. Their amount was evaluated by flow cytometry analysis. The proportion of labeled (apoptotic) cells was measured by flow cytometry analysis relative to negative controls labeled with dUTP without terminal deoxynucleotidyl transferase.

Charrad, R. S., Gadhoum, Z., Qi, J., Glachant, A., Allouche, M., Jasmin, C.,... & Smadja-Joffe, F. (2002). Effects of anti-CD44 monoclonal antibodies on differentiation and apoptosis of human myeloid leukemia cell lines. Blood, 99(1), 290-301.

Figure 4 A3D8 induces monocytic differentiation of THP-1 and HL60 cells.

The data have been determined after addition of 2.5 μg/mL A3D8 at day 5 in the case of THP-1 cells.

Charrad, R. S., Gadhoum, Z., Qi, J., Glachant, A., Allouche, M., Jasmin, C.,... & Smadja-Joffe, F. (2002). Effects of anti-CD44 monoclonal antibodies on differentiation and apoptosis of human myeloid leukemia cell lines. Blood, 99(1), 290-302.

Figure 5 A3D8 induces monocytic differentiation of THP-1 and HL60 cells.

Induction of synthesis of IL-1β, G-CSF, and IL-3 transcripts in HL60 cells. Total RNA from control and treated HL60 cells was prepared as described in Materials and methods. Using primers for glyceraldehyde phosphodehydrogenase, the amounts of cDNA were equilibrated to this internal standard. RT-PCR amplification products were separated by 1% agarose gel electrophoresis with 0.5 μg/mL ethidium bromide and were pictured under UV-light irradiation.

Charrad, R. S., Gadhoum, Z., Qi, J., Glachant, A., Allouche, M., Jasmin, C.,... & Smadja-Joffe, F. (2002). Effects of anti-CD44 monoclonal antibodies on differentiation and apoptosis of human myeloid leukemia cell lines. Blood, 99(1), 290-303.