human recombinant mgmt  (Cayman Chemical)

Journal: PLoS ONE

Article Title: Fluorogenic Real-Time Reporters of DNA Repair by MGMT, a Clinical Predictor of Antitumor Drug Response

doi: 10.1371/journal.pone.0152684

Figure Lengend Snippet: Fluorescence spectra showing the overall fold-changes in intensity observed when comparing fluorescence measured before (dashed line) and after (solid line) addition of purified MGMT protein are shown at left of each figure. Time courses (on the right) show time-dependent fluorescence increases immediately after addition of enzyme. Final probe and MGMT concentrations were 100 nM. Assays were run at 37°C in 70 mM HEPES buffer pH 7.8 containing 5 mM EDTA, 1 mM dithiothreitol and 50 μg/ml BSA. (A) chemosensor 1 containing dT FAM , (B), chemosensor 2 containing Cy3, (C), chemosensor 3 containing dT TMR and (D), chemosensor 4 containing perylene nucleoside. Measurements were repeated 3 times. Standard deviations are provided in .

Article Snippet: Purified recombinant human MGMT (His-tagged, expressed in E . Coli ) was purchased from Creative BioMart.

Techniques: Fluorescence, Purification

Journal: PLoS ONE

Article Title: Fluorogenic Real-Time Reporters of DNA Repair by MGMT, a Clinical Predictor of Antitumor Drug Response

doi: 10.1371/journal.pone.0152684

Figure Lengend Snippet: Incubation of purified MGMT enzyme with the inhibitors BG and PaTrin-2 led to a concentration dependent decrease in observed final fluorescence intensity, indicative of MGMT inhibition. MGMT (10 nM) was incubated with inhibitor for 10 min at 37°C in 70 mM HEPES buffer (pH 7.8) containing 5 mM EDTA, 1 mM dithiothreitol and 50 μg/ml BSA. Final fluorescence was acquired 10 min after addition of probe (10 nM). Data were normalized to measurements without inhibitor. Each data point is the average of 3 measurements.

Article Snippet: Purified recombinant human MGMT (His-tagged, expressed in E . Coli ) was purchased from Creative BioMart.

Techniques: Incubation, Purification, Concentration Assay, Fluorescence, Inhibition

Journal: Veterinary Sciences

Article Title: Alkylation and Carbamylation Effects of Lomustine and Its Major Metabolites and MGMT Expression in Canine Cells

doi: 10.3390/vetsci2020052

Figure Lengend Snippet: Graph showing MGMT mRNA expression in 17–71 cells before (untreated) and after treatment with lomustine at 1000, 3000 and 12,000 ng/mL and incubated for 6, 12, 24 and 48 h.

Article Snippet: In order to ascertain the functionality of the assay, human recombinant MGMT (Cayman chemicals) was used as a positive control.

Techniques: Expressing, Incubation

Journal: Veterinary Sciences

Article Title: Alkylation and Carbamylation Effects of Lomustine and Its Major Metabolites and MGMT Expression in Canine Cells

doi: 10.3390/vetsci2020052

Figure Lengend Snippet: Graph showing MGMT mRNA expression in 17–71 cells before (untreated) and after treatment with trans-4-hydroxylomustine and cis-4 hydroxylomustine at 12,000 ng/mL concentration and incubated for 6, 12, 24 and 48 h.

Article Snippet: In order to ascertain the functionality of the assay, human recombinant MGMT (Cayman chemicals) was used as a positive control.

Techniques: Expressing, Concentration Assay, Incubation

Journal: Veterinary Sciences

Article Title: Alkylation and Carbamylation Effects of Lomustine and Its Major Metabolites and MGMT Expression in Canine Cells

doi: 10.3390/vetsci2020052

Figure Lengend Snippet: Graphs showing the MGMT mRNA expression in canine lymphoma cell line 17–71 cells and primary hepatocytes from six dogs.

Article Snippet: In order to ascertain the functionality of the assay, human recombinant MGMT (Cayman chemicals) was used as a positive control.

Techniques: Expressing

Journal: Veterinary Sciences

Article Title: Alkylation and Carbamylation Effects of Lomustine and Its Major Metabolites and MGMT Expression in Canine Cells

doi: 10.3390/vetsci2020052

Figure Lengend Snippet: ( A ) MGMT activity in canine lymphoma cell line 17–71: lanes 1–3 represent replicate oligo samples treated with 500 μg cell protein from 17–71 cells. Lanes labeled “N” are untreated oligo samples (control). Upper and lower bands represent 18 bp substrate and 10 bp cleavage product, respectively; ( B ) MGMT activity in primary hepatocytes obtained from six dogs: lanes 1–6 represent oligo samples treated with 500 μg cell protein from hepatocytes obtained from six dogs. Lanes labeled “N” are untreated oligo samples (control). Upper and lower bands represent 18 bp substrate and 10 bp cleavage product, respectively.

Article Snippet: In order to ascertain the functionality of the assay, human recombinant MGMT (Cayman chemicals) was used as a positive control.

Techniques: Activity Assay, Labeling, Control

Journal: Veterinary Sciences

Article Title: Alkylation and Carbamylation Effects of Lomustine and Its Major Metabolites and MGMT Expression in Canine Cells

doi: 10.3390/vetsci2020052

Figure Lengend Snippet: Graph showing MGMT enzyme activity in the primary hepatocytes obtained from six dogs and in the canine lymphoma cell line 17–71.

Article Snippet: In order to ascertain the functionality of the assay, human recombinant MGMT (Cayman chemicals) was used as a positive control.

Techniques: Activity Assay

Journal: Veterinary Sciences

Article Title: Alkylation and Carbamylation Effects of Lomustine and Its Major Metabolites and MGMT Expression in Canine Cells

doi: 10.3390/vetsci2020052

Figure Lengend Snippet: Graph showing correlation between MGMT gene expression and enzyme activity in canine lymphoma cell line 17–71 and the hepatocytes obtained from six dogs. Pearson’s correlation was 0.806 which was highly significant with p value of 0.028. ( p value set at 0.05).

Article Snippet: In order to ascertain the functionality of the assay, human recombinant MGMT (Cayman chemicals) was used as a positive control.

Techniques: Gene Expression, Activity Assay