MUC5AC Monoclonal Antibody (45M1)

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	-Mouse (1:500)

Protocol tips
-Mouse (1:500)

Publication protocol

Immunoblotting for MUC5AC was carried out under nonreducing conditions as described by Lacunza and colleagues (45). Briefly, BAL and ASL samples were diluted in SDS-PAGE sample buffer containing 25% SDS (125 mM Tris·HCl, pH 6.8; 25% SDS; 20% glycerol; 0.004% bromophenol blue) prior to boiling at 95°C. Samples were loaded onto 7.5% Tris·HCl gels (5 μg/lane) and electrophoresed for ∼2 h. Then proteins were transferred to PVDF membranes by electroblotting overnight at 4°C, by using methanol-free transfer buffer containing 0.1% SDS. To block nonspecific immunoreactivity, the membranes were incubated for 1–2 h at room temperature in TBS-Tween containing 2% BSA. Then they were incubated for 1.5 h with a monoclonal antibody recognizing MUC5AC (Clone 45M1; Thermo Fisher Scientific) diluted 1:500 in TBS-Tween. Next they were washed four times in TBS-Tween and incubated for 1 h with secondary antibody (Immunopure goat anti-mouse conjugated to horseradish peroxidase; Thermo Fisher Scientific) at a 1:20,000 dilution. After five washes in TBS-Tween, bands were detected as described above for PLUNC. Electrophoresis and immunoblotting of MUC5AC under these conditions is expected to produce a predominant band of ∼200 kDa

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for MUC5AC Monoclonal Antibody (45M1) below.

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