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The reconstituted HqD complex showing positive HqD activity (described later) was determined by comparing the elution volume in the gel filtration chromatography and its comparison with standard proteins with known molecular weights.
Calibration curve using standard proteins with known sedimentation coefficients and linear regression curve (E ).
The void volume of the column (V0) and the elution position of standard proteins with their respective stokes radii is indicated.
To demonstrate the effectiveness of pMatch, a library search experiment was conducted on a public dataset of standard proteins with over 40 000 spectra.
Accordingly, we assessed its HIC performance via individual injections of the same set of 10 standard proteins with ammonium tartrate as the salt.
Furthermore, it is guaranteed that any possible peptide resulting from digesting the seven standard proteins with trypsin is not present in the random database.
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For quantification of relative signal intensities, each Western blot contained a dilution series of a standard protein with known concentration.
Assays were performed in duplicate, and readings were compared with standard curves obtained with standard protein provided with the kit.
Subsequently, total protein content was measured with the bicinchinonic acid system (Uptima, France) using bovine serum albumin as standard, proteins were reduced with 2-mercaptoethanol and total protein concentrations adjusted.
The pI of the isoforms were estimated by comparison to standard protein of known pI value run on the same IEF gel with the track of the standard proteins separated and visualised with Coomassie blue protein stain rather than with AP zymography.
Apparent molecular weights were determined using a calibration plot with standard proteins as a reference (broad range SDS-PAGE Standards, Bio-Rad Laboratories). Bio-Rad Laboratories
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