leupeptin

DTT, phosphatase inhibitors (0.1 µM okadaic acid and 1 mM sodium orthovanadate) and protease inhibitors (1 mM PMSF and 10 µg/ml leupeptin) using a motorized teflon-glass homogenizer.

PLoS ONE Alerts: New Articles Pramod K. Dash et al. 2010

Tissues were homogenized in a lysis buffer consisting of 10 mM Hepes, 1% Triton X-100, and protease inhibitor cocktail, including aprotinin, leupeptin, pepstatin A, Bestatin, E-64,

PLoS ONE Alerts: New Articles Franco Folli et al. 2010

To examine the functional consequences of the blockade of calpain activity on the UCB effects, slices were co-exposure to UCB (10 µM) and leupeptin or calpeptin.

PLoS ONE Alerts: New Articles Fang-Yu Chang et al. 2009

The diluting buffer contained the same amounts of DTT and leupeptin but only 0.2 mM PMSF.

PLoS ONE Alerts: New Articles 2009

HaCaT cells were lysed in 50 mM Tris-Cl, pH 7.4 with 5 mM EDTA, 250 mM NaCl, and 0. 1% Triton X-100 (lysis buffer) plus protease inhibitors (1 mM PMSF, 10 µg/ml aprotinin and leupeptin) for 30-40 min on ice.

PLoS ONE Alerts: New Articles 2009

(A) Representative immunoblots showing protein levels of NR1, NR2A, and NR2B subunits in slices treated with 10 µM UCB for 48 h in the absence or presence of calpain inhibitors, leupeptin (100 µM) or calpeptin (100 µM).

PLoS ONE Alerts: New Articles Fang-Yu Chang et al. 2009

Cells were washed 3 times in PBS, and detached from the plate surface with cell scrapers in the presence of protease inhibitors (sodium pyrophosphate, sodium fluoride, phenylmethanesulfonyl fluoride, pepstatin A, aprotinin, leupeptin, antipain, benzamidine - all from Sigma) diluted in PBS.

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DTT (0.5 mM), PMSF (0.5 mM), and leupeptin (10 pg / ml) were added to the lysis and extraction buffers just before use.

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Cell extracts were obtained with RIPA buffer in the presence of protease inhibitors (sodium pyrophosphate, sodium fluoride, phenylmethanesulfonyl fluoride, pepstatin A, aprotinin, leupeptin, antipain, benzamidine - all from Sigma).

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Figure 10A-D, both leupeptin (100 µM) and calpeptin (100 µM) completely prevented UCB-induced down-regulation of NR2A and NR2B subunit proteins.

PLoS ONE Alerts: New Articles Fang-Yu Chang et al. 2009