Mouse Anti-Carbohydrate Recombinant Antibody (clone BR96); Fab Fragment (CAT#: PFBC-018)

Figure 1 BR96 cell surface localization.

Fixed 3396 ceils were stained with BR96/gold and viewed by darkfield microscopy. Bright spots represent focal accumulations of BR96 (arrowheads). Unlabeled membrane regions are dark green/blue (Fig. 1 a, open arrow). At the cell margin, BR96 labeled ruffled membranes are pink/red (arrows). BR96 is expressed at the leading lamella of migrating cells (Fig. 1 c, open arrow). Bar, 25 μm.

Garrigues, J., Anderson, J., Hellström, K. E., & Hellström, I. (1994). Anti-tumor antibody BR96 blocks cell migration and binds to a lysosomal membrane glycoprotein on cell surface microspikes and ruffled membranes. The Journal of cell biology, 125(1), 129-142.

Figure 2 BR96 localization using permeabilized cells.

184 mammary epithelial cells were fixed, permeabilized with 'Triton X-100, and stained with BR96/FITC. BR96 localized to ruffled membranes at the cell margin (open arrow), microspikes (arrowheads), and to vesicles (arrows) found throughout the cytoplasm especially in the perinuclear region. Bar, 20 μm.

Garrigues, J., Anderson, J., Hellström, K. E., & Hellström, I. (1994). Anti-tumor antibody BR96 blocks cell migration and binds to a lysosomal membrane glycoprotein on cell surface microspikes and ruffled membranes. The Journal of cell biology, 125(1), 129-142.

Figure 3 BR96 inhibits cell migration.

Cultures of 3396 carcinoma cells were scratch wounded and treated with either BR96 (a) or IIG5 control (b) for 65 h. The cells were fixed and stained with Dapi. The white line marks the location of the wound edge just before mAb treatment. The arrows in a indicate areas of cell aggregation.

Garrigues, J., Anderson, J., Hellström, K. E., & Hellström, I. (1994). Anti-tumor antibody BR96 blocks cell migration and binds to a lysosomal membrane glycoprotein on cell surface microspikes and ruffled membranes. The Journal of cell biology, 125(1), 129-142.

Figure 4 BR96 inhibits cell migration.

The percent migration was calculated by measuring the change in area of the scratch which remained cell free after treatment with BR96 (solid line) or fIG5 (dashed line) as compared to media control (c). In some experiments after BR96 treatment, the cells were cultured further (24 h) in 15 % IMDM without mAb and stained with BR96/gold (d, montage). Note the prominent microspikes extending from the cell margin are heavily labeled with BR96/gold which appears dark brown; adjacent membrane regions (d, open arrows) are stained significantly less.

Garrigues, J., Anderson, J., Hellström, K. E., & Hellström, I. (1994). Anti-tumor antibody BR96 blocks cell migration and binds to a lysosomal membrane glycoprotein on cell surface microspikes and ruffled membranes. The Journal of cell biology, 125(1), 129-142.

Figure 5 Immunoblot

BR96 antigens were purified on 6 % tris glycine preparative gels, eleetroeluted, and aliquots of the purified protein were either iodinated or blotted and probed with BR96 (c).

Garrigues, J., Anderson, J., Hellström, K. E., & Hellström, I. (1994). Anti-tumor antibody BR96 blocks cell migration and binds to a lysosomal membrane glycoprotein on cell surface microspikes and ruffled membranes. The Journal of cell biology, 125(1), 129-142.