... engineered in place of the FLAG tag [25] Site-directed mutagenesis was carried out using the QuikChangeÔ mutagenesis kit from Stratagene (La Jolla, CA, USA) Pfu-directed thermal cycling was conducted ... Man-6-P to compensate for nonspecific binding The data were graphed and analyzed using graphpad prismÔ (Graphpad Software, San Diego, CA, USA) Ligand binding parameters were measured by competitive ... above The data were fit to a model for one-site and two-site competitive binding using graphpad prismÔ Acknowledgements We thank Dr William S Sly for providing the human M6P ⁄ IGF2R cDNA, Dr David...
... rabbit polyclonal (Santa Cruz Biotechnology, Santa Cruz, CA, USA), directly labeled using DyLightÔ549 Microscale Antibody Labeling Kit (Pierce, Rockford, IL, USA); antiMUPP1, rabbit polyclonal ... free from artificial odorant stimulation All mice were held in 7288 Peptide microarray CelluSpotsÔ Peptide Arrays (Intavis AG, Cologne, Germany) were blocked for h at room temperature with 5% skimmed ... incubated with a PDZ1 + 2_HA fusion protein (produced as described in E coli) overnight at °C CelluSpotsÔ were incubated with the a-HA antibody for h at room temperature Detection was performed with HRP...
... the enzyme is active only with the ÔcorrectÕ metal Although, in the complete absence of the ÔcorrectÕ metal, each type of SOD may be forced to fold with the ÔincorrectÕ metal at its active centre, ... of each mutant enzyme on a Ôper metal ionÕ basis using values obtained for the correct metal Relative results not vary significantly from the specific activities on a Ôper mg proteinÕ basis, except ... During catalysis substrate and products must pass through a ÔfunnelÕ made up of residues from each subunit [11,24] and include so-called Ôgateway residuesÕ His30, Tyr34, Trp169 and Glu170, the latter...