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4 protocols using mmp 2 mmp 3 inhibitor 1

1

Antibodies and Reagents for β1-Adrenergic Receptor Analysis

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Antibodies were from the following sources: rabbit polyclonal anti-β1AR (Cat# ab3442, raised against residues 394-408 in human β1-ARs) was from Abcam. Mouse monoclonal anti-FLAG M2 antibody (Cat# F1804) was from Sigma-Aldrich. The Abcam rabbit polyclonal anti-β1AR was validated in our previous publication.3 IRDye 800CW goat anti-rabbit IgG (Cat# 925-32211) and IRDye 680RD goat anti-mouse IgG (Cat# 925-68070) secondary antibodies were from LI-COR Biosciences. Elastase from porcine pancreas (Cat# E0258), GM6001 (Cat# 364206), GI254023X (ADAM-10 inhibitor, Cat# SML0789), matrix metalloproteinase (MMP)-2/MMP-9 inhibitor I (Cat# 444241), MMP-9/MMP-13 inhibitor I (Cat# 444252), forskolin (Cat# F3917), isoproterenol (Iso) (Cat# 420355), propranolol (Cat# P0884), sotalol (Cat# S0278), and theophylline (Cat# T1633) were from Sigma-Aldrich. MMP-2/MMP-3 inhibitor I (Cat# sc-295483) and TAPI-2 (Cat# sc-205851) were from Santa Cruz Biotechnology. GW280264X (ADAM-10/ADAM-17 inhibitor, Cat# 7030) was from AOBIOUS Inc. α2-3,6,8,9 neuraminidase A (Cat #: P0722) was from New England BioLabs Inc. All other chemicals were reagent grade.
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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

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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4

Antibody Sources and Inhibitor Reagents

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Antibodies were from the following sources: Rabbit polyclonal anti-βAR (ab3442, raised against residues 394-408 in human β1AR) was from Abcam (Cambridge, MA). Mouse monoclonal anti-FLAG M2 and ant-β-actin were from Sigma-Aldrich (Saint Louis, MO). Rabbit monoclonal anti-cleaved caspase-3 (Asp175) (5A1E) was from Cell Signaling Technology (Danvers, MA). IRDye 800CW or 680RD goat anti-rabbit or goat anti-mouse IgG (H+L) were from LI-COR Biosciences (Lincoln, NE). GM6001, GI254023X (ADAM-10 inhibitor), MMP-2/MMP-9 inhibitor II, MMP-9/MMP-13 inhibitor II, doxorubicin, and H2O2 were from Sigma-Aldrich. MMP-2/MMP-3 inhibitor I and TAPI-2 were from Santa Cruz Biotechnology (Dallas, TX). GW280264X (ADAM-10/ADAM-17 inhibitor) was from AOBIOUS Inc (Hopkinton, MA). All other chemicals were reagent grade.
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