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Other chemicals and reagents used in this study were of analytical grade. Plasmids and lentiviruses ATP6AP2-V5-FL and ATP6AP2-V5-ICD plasmids were purchased from DNASU (ATP6AP2 in pLX304, HsCD00446844 and HsCD00437681). ATP6AP2-V5-ECD was made by cloning the ECD domain (17-274) ...
The cleaved protein was passed through Ni-NTA resin to remove the 6xHis-GST fusion protein and TEV protease prior to size exclusion chromatography on a prep-grade Superdex S200 (Cytiva) column in 50 mM Tris pH 8.0, 500 mM NaCl, 1 mM TCEP, and 5% glycerol. Fractions containing ...
For DNA digestion of single and double-stranded DNA, 1 μg RNA sample was incubated at room temperature for 10 min with 1 μl 10× DNase I Reaction Buffer, 1 μl DNase I Amp Grade 1 U/μl, and DEPC-treated water to final volume of 10 μl. DNase I was inactivated by incubation ...
Briefly, 100,000 cells (parental THP-1 cells and 7CKO clone B11, 3 independent experiments each) were centrifuged, and resuspended in 500 µL of cold 0.9% NaCl. Metabolites were extracted with the addition of 600 µL LC/MS-grade cold methanol containing internal standards, and the...
Complete answer:Single-celled microorganisms known as prokaryotic cells are the oldest on the planet....
The proteins were digested by adding a solution of trypsin in 50 mM NH4HCO3 (20 ng/μl sequence grade, Promega). The mixture was incubated for 3 h at 37 °C. 2.18. Nanoelectrospray tandem mass spectrometry Recovered peptides were subjected to nanoLC-MS/MS analysis on an ESI-Q-TOF mass...
An analysis was performed to determine the correlation between T2 MRI values in endometrial cancer (EC) and pathological characteristics, including International Federation of Gynecology and Obstetrics (FIGO) stage and grade.Middling T2 values for NE, BEL, and EC were found to be 1975 ms (ranging...
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Adenosine triphosphate (ATP) plays fundamental roles in cellular biochemistry and was recently discovered to function as a biological hydrotrope. Here, we use mass spectrometry to interrogate ATP-mediated regulation of protein thermal stability and prote