Bml zw9345 0005

Manufactured by Enzo Life Sciences

BML-ZW9345-0005 is a laboratory equipment product from Enzo Life Sciences. It is designed for specific laboratory applications. A detailed description of the product's core function is not available while maintaining an unbiased and factual approach.

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3 protocols using bml zw9345 0005

Proteasome Activity Assay Protocol

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Proteasome activity assays were performed as described previously [31 (link), 45 , 46 (link)]. Normalized samples (nuclear = 2μg; cytoplasmic = 20μg) were diluted in MilliQ H₂O and mixed with reaction buffer (250mM HEPES, pH 7.5, 5mM EDTA, 0.5% NP-40, 0.01% SDS, 5mM ATP). Fluorogenic peptides Suc-leu-leu-val-thy-AMC (#BML-P802-0005, Enzo Life Sciences), Bz-val-gly-arg-AMC (#BML-BW9375-0005, Enzo Life Sciences) and z-leu-leu-glu-AMC (#BML-ZW9345-0005; Enzo Life Sciences) were then added to the samples at a concentration of 10μM to assess proteasome chymotrypsin-like, trypsin-like and peptidylglutamyl-peptide hydrolyzing-like activities, respectively. The reaction was incubated at 37°C for 2 hours and fluorescence monitored at 360 (excitation)/460 (emission) on a monochromatic plate reader (Synergy H1; Biotek, Winooski, VT). Protein free blanks were used and an AMC standard curve was produced. The scan with the peak level of AMC was used for statistical analyses. Data are presented as the percentage change in relative fluorescent units (RFU) relative to the Control group.

McFadden T., Musaus M., Nelsen J.L., Martin K., Jones N., Smith P., Kugler H, & Jarome T.J. (2020). Dysregulation of protein degradation in the hippocampus is associated with impaired spatial memory during the development of obesity. Behavioural brain research, 393, 112787.

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Proteasome Activity Assays

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Proteasome activity assays were performed as described previously [15 (link), 28 ]. Normalized samples (nuclear = 2μg; cytoplasmic = 20μg) were diluted in MilliQ H₂O and mixed with reaction buffer (250mM HEPES, pH 7.5, 5mM EDTA, 0.5% NP-40, 0.01% SDS, 5mM ATP). Fluorogenic peptides Suc-leu-leu-val-thy-AMC (#BML-P802-0005, Enzo Life Sciences), Bz-val-gly-arg-AMC (#BML-BW9375-0005, Enzo Life Sciences) and z-leu-leu-glu-AMC (#BML-ZW9345-0005; Enzo Life Sciences) were then added to the samples at a concentration of 10μM to assess proteasome chymotrypsin-like, trypsin-like and peptidylglutamyl-peptide hydrolyzing-like activities, respectively. The reaction was incubated at 37°C for 2 hours and fluorescence monitored at 360 (excitation)/460 (emission) on a monochromatic plate reader (Synergy H1; Biotek, Winooski, VT). Protein free blanks were used and an AMC standard curve was produced. The scan with the peak level of AMC was used for statistical analyses. Data are presented as the percentage change in relative fluorescent units (RFU) relative to the Control group.

McFadden T., Farrell K., Martin K., Musaus M, & Jarome T.J. (2022). Short-term exposure to an obesogenic diet causes dynamic dysregulation of proteasome-mediated protein degradation in the hypothalamus of female rats. Nutritional neuroscience, 26(4), 290-302.

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Proteasome Activity Assay Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Proteasome activity assays were performed as described previously [15, 28] . Normalized samples (nuclear = 2μg; cytoplasmic = 20μg) were diluted in MilliQ H 2 O and mixed with reaction buffer (250mM HEPES, pH 7.5, 5mM EDTA, 0.5% NP-40, 0.01% SDS, 5mM ATP). Fluorogenic peptides Suc-leu-leu-val-thy-AMC (#BML-P802-0005, Enzo Life Sciences), Bz-val-gly-arg-AMC (#BML-BW9375-0005, Enzo Life Sciences) and z-leuleu-glu-AMC (#BML-ZW9345-0005; Enzo Life Sciences) were then added to the samples at a concentration of 10μM to assess proteasome chymotrypsin-like, trypsin-like and peptidylglutamyl-peptide hydrolyzing-like activities, respectively. The reaction was incubated at 37°C for 2 hours and fluorescence monitored at 360 (excitation)/460 (emission) on a monochromatic plate reader (Synergy H1; Biotek, Winooski, VT). Protein free blanks were used and an AMC standard curve was produced. The scan with the peak level of AMC was used for statistical analyses. Data are presented as the percentage change in relative fluorescent units (RFU) relative to the Control group.

McFadden T., Farrell K., Martin K., Musaus M, & Jarome T.J. (2023). Short-term exposure to an obesogenic diet causes dynamic dysregulation of proteasome-mediated protein degradation in the hypothalamus of female rats. Nutritional neuroscience, 26(4).

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