25 mM TrisHCl, pH7.5, 100 mM KCl, 5 mM MgCl2, 10% (v/v) glycerol. Protein concentration was determined with a BCA assay-kit from Invitrogen using bovine serum albumin for the calibration curve. GTP Binding Analysis: Fluorescent emission spectra of FA/AF11A were obtained at ˜1 μM in...
The double stand DNA was inserted into pENTR/ H1/TO vector to generate the pENTR/H1/TO-Bcl- xL siRNA construct using BLOCK-iTTM Inducible H1 RNAi Entry Vector Kit (Invitrogen, Carlsbad, CA). Transfection of pENTR/H1/TO-Bcl-xL siRNA was performed by electroporation using a Gene Pulser II...
Fast PCR software was used to design primer fromCtenopharyngdon idellaandDanio rerioMx to get a full-length sequence of Mx cDNA. Seven set of primer was designed to amplify different segments of the Mx gene (See Table3). The amplified product was purified, using PCR purification kit (Gen ...
Precipitated materials were removed by centrifugation, and the IP3 content of the supernatant was measured by a competitive radioreceptor assay using an IP3 3H Radioreceptor Assay Kit (NEN). 3. Results 3.1. Cloning and characterization of cDNAs for Gαq family proteins Initially, we conducted PCR...
RevertAid Reverse Transcriptase kit ThermoFisher Cat#K1691 Deposited data Proteomics: Raw spectrum runs, supporting worked up raw files and metadata this study MassIVE: MSV000088135 Experimental models: Organisms/strains S. cerevisiae: BY4742 (referred to as “WT”):MATα his3Δ1 leu2Δ0 lys2Δ...
GTPase Colorimetric Assay Kit 480 Tests Innova Biosciences Cat# 602-0121 Deposited Data Gαs(R201C/C237S) structure This paper PDB: 6AU6 Experimental Models: Cell Lines Sf9 cells Thermo Fisher Scientific Cat# 12659017 Recombinant DNA MGC Human ADCY2 Sequence-Verified cDNA GE Healthcare Life Sci...
To investigate the significance of the NOG1-2 and JAZ9 interaction, we assessed the GTPase activity of recombinant purified NOG1-2, in the presence of different concentrations of JAZ9, in a real time fluorescence-based GTP-binding/hydrolysis and a phosphate release assay [24,25] (Figure 3...