To these solution add Glucose (10mM; 0.4507g/250ml) This buffer is made the same as the 1.8mM Ca2+ buffer, however the amount of Ca2+ added is different.
It was then incubated for 1.5 h with 0.5 μM of either of the hexahistidine-tagged 7-, 6-, or 5-repeat DC-SIGNR variants in buffer A. Nonspecifically adsorbed protein was removed by rinsing the bilayer with buffer A. The amount of DC-SIGNR immobilized on the supported NTA-TRIG-DLGE monolayers was quantified by surface plasmon resonance.
The carbonate buffer decreased the amount of ordered structure in starch as judged by DSC enthalpy values, while increasing the melting temperature of these structures.
After 48 h, samples were lysed using 1x Passive lysis buffer, and the amount of firefly luciferase and Renilla luciferase was quantified using the Dual-Luciferase Reporter Assay System Promega Corporationn, Masison, WI), as we described recently [15].
The method can be adapted to each species by modification of the digestion buffer, of the amount of the enzymatic cocktail added during the digestion or of the digestion time.
Colospheres, spheroids and xenograft tissue were lysed with zymogram sample buffer and the amount of protein was estimated using the Bradford method (Biorad Dc Protein Assay; Bio-Rad Laboratories).
This indicates that ferritin acts indeed as a buffer, keeping the amount of iron in the cLIP essentially independent from the iron-to-ferritin ratio.
Protein bound to the beads was washed three times in lysis buffer and the amount of bound fusion protein was estimated using Coomassie-stained SDS gels.
After immunoprecipitation, beads were washed three times with lysis buffer and the amount of HCF101 Myc-associated radioactivity was measured by scintillation counting.
Tissue samples or myofibroblasts were homogenized in ice-cold lysis buffer, and the amount of protein was determined by a protein assay (Bio-Rad Laboratories).
