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4 protocols using s2672

1

Targeted Metalloproteinase Inhibition

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Batimastat at 20nM (Pan MMP inhibitor; BB-94; Selleck Chemicals); MMP-2/MMP-9 Inhibitor II at 50nM (sc-311430; Santa Cruz biotechnology); ARP 100 at 20nM (MMP2 inhibitor) (R&D Systems); MMP-13 Inhibitor at 10nM (sc-205756; Santa Cruz biotechnology); MMP-9 Inhibitor I at 100nM (sc-311437; Santa Cruz biotechnology); MMP-9/MMP-13 Inhibitor II at 10nM (sc-311439; Santa Cruz biotechnology); MMP2/MMP-3 Inhibitor I at 20uM (sc-295483; Santa Cruz biotechnology); TAPI-1 at 20uM (ADAM17 inhibitor; S7434; Selleck Chemicals); GI254023X at 200nM unless otherwise indicated (ADAM10 inhibitor; Sigma Aldrich); PF-00562271 at 5nM (FAK inhibitor; S2672; Selleck Chemicals).
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2

Targeted Metalloproteinase Inhibition

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Batimastat at 20nM (Pan MMP inhibitor; BB-94; Selleck Chemicals); MMP-2/MMP-9 Inhibitor II at 50nM (sc-311430; Santa Cruz biotechnology); ARP 100 at 20nM (MMP2 inhibitor) (R&D Systems); MMP-13 Inhibitor at 10nM (sc-205756; Santa Cruz biotechnology); MMP-9 Inhibitor I at 100nM (sc-311437; Santa Cruz biotechnology); MMP-9/MMP-13 Inhibitor II at 10nM (sc-311439; Santa Cruz biotechnology); MMP2/MMP-3 Inhibitor I at 20uM (sc-295483; Santa Cruz biotechnology); TAPI-1 at 20uM (ADAM17 inhibitor; S7434; Selleck Chemicals); GI254023X at 200nM unless otherwise indicated (ADAM10 inhibitor; Sigma Aldrich); PF-00562271 at 5nM (FAK inhibitor; S2672; Selleck Chemicals).
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3

Comprehensive Antibodies and Reagents Protocol

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Total β-catenin antibody (1:1,000) was purchased from Invitrogen (712700), glyceraldehyde-3-phosphate dehydrogenase antibody (1:1,000) and FAK antibody (1:1,000, 3285) were obtained from Cell Signaling Technologies, anti-ATP6V0a3 antibody (1:500) was obtained from Novus (nbp1–89333, 1:1,000). CD63 antibody was obtained from Abcam.
Antibodies against Pak1 (ab131522), Ras (ab52939), GSK3 (ab93926, 1:4000), and secondary antibodies for immunostaining for cells (ab150120, ab150084, ab150117, ab150081) (1:300) were obtained from Abcam. Antibody against the focal adhesion protein Tes (HPA018123, 1:100) was obtained from Atlas antibodies. Secondary antibodies for immunostaining arrays (A-11001, A-11011 were obtained from Invitrogen). HRP-linked secondary antibodies (7076, 7074 at 1:5000 (Cell Signaling) were used for Western blots and analyzed with an iBright Imaging system. EIPA (A3085), and LiCl (L4408), were obtained from Sigma. Baf (S1413) and PF-00562271 (FAK inhibitor, S2672) were purchased from Selleckchem. TMR- dextran 70 kDa was obtained from ThermoFisher (D1818). PMA (1201) was purchased from TOCRIS.
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4

Cell Signaling Pathway Antibody Protocol

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Total β-catenin antibody (1:1000) was purchased from Invitrogen (712700), glyceraldehyde-3-phosphate dehydrogenase antibody (1:1000) and FAK antibody (1:1000, 3285) were obtained from Cell Signaling Technologies, anti-ATP6V0a3 antibody (1:500) was obtained from Novus (nbp1-89333, 1:1000). CD63 antibody was obtained from Abcam.
Antibodies against Pak1 (ab131522), Ras (ab52939), GSK3 (ab93926, 1:4000), and secondary antibodies for immunostaining for cells (ab150120, ab150084, ab150117, ab150081) (1:300) were obtained from Abcam. Antibody against the FA protein Tes (HPA018123, 1:100) was obtained from Atlas antibodies. Secondary antibodies for immunostaining arrays (A-11001, A-11011 were obtained from Invitrogen). HRP-linked secondary antibodies 7076, 7074 at 1:5000 (Cell Signaling) were used for western blots and analyzed with an iBright Imaging system. EIPA (A3085), and LiCl (L4408), were obtained from Sigma. Baf (S1413) and PF-00562271 (FAK inhibitor, S2672) were purchased from Selleckchem. TMR-dextran 70 kDa was obtained from Thermo Fisher (D1818). PMA (1201) was purchased from TOCRIS.
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