1. B. MAb 9E1 reduces MMP-9 activity in MDA-MB-231 cells. A
Figure 1: Characterisation of MAb 9E1.
A. MAb 9E1 reacts with a 65 kDa reactive band
A 65Kda reactive band is identified across breast cancer cell lines. BT-20 does not express the MAb target antigen and reduced expression is observed in MDA- MB-468 (absent in membrane enriched fractions). Reactivity is observed in both total cell lysates and membrane enriched fractions. Duplicate biological samples were probed with MAb 9E1, a representative immunoblot is shown.
B. MAb 9E1 reduces MMP-9 activity in MDA-MB-231 cells.
(i) Gelatin zymography of conditioned medium from MDA-MB-231 cells with intense clear zone showing active MMP-9 (92 kDa). A very weak signal indicative of pro MMP-9 can be observed directly underneath the active form.
(ii) Representative gelatin zymogram showing analysis of conditioned medium (n=2). following treatment of cells with 50ug/ml of MAb 9E1. A reduction in MMP-9 activity is observed in antibody treated conditioned medium compared relative to control hybridoma medium only (no antibody). Densitometry analysis of reduced MMP-9 activity (fold change) is also shown. . A
Whole cell lysates
Membrane enriched fractions
Alpha tubulin B i ii
110 kDa
80 kDa
active MMP-9
110 KDa
active MMP-9
Fold change
Hybridoma medium control MAb 9E1
(no antibody)