After a 10-min incubation, the suspension was centrifuged at 16,000 rpm for 15 min at 4°C to remove the cellular debris, and the membrane vesicles were then harvested by centrifugation at 125,000 rpm at 4°C for 1 h, and resuspended in 1.5 ml buffer A (50 mM Hepes pH 8.0, 250 mM NaCl, 10% glycerol [v/v]), and used as the protein sample in immediate assays for ATPase activity [ 1].
Immediate assay of the samples in the presence of reconstituted milk powder was essential for luciferase detection in female guts.
The venous blood samples were collected in 10-ml vacutainers containing lithium-heparin (Becton Dickinson), which were placed on ice and transported to our laboratory for immediate assay.
The venous blood samples were collected in 10-ml vacutainers containing lithium-heparin (Becton Dickinson Biosciences, San Jose, CA, USA), which were placed on ice and transported to our laboratory for immediate assay.
Compounds were chosen based upon structural comparison to the compounds 1 and 2, and ready availability either from commercial sources for immediate assay, or by Knoevenagel condensation from precursor aldehydes.
Blood and urine specimens will be taken for immediate routine assays as well as for freezing pending peptide and genetic studies.
The abundant SNPs can be used to design more targeted SNP assays for immediate use in rice genetics and molecular breeding [ 8- 10], particularly in Genome-wide association study (GWAS) [ 11- 13].
and assayed the immediate molecular responses to wounding in wild-type and mutant corneal epithelial cells.
Interestingly, the maximal increase in the percentage of RFP+SST+ cells was evident by 2DIV, suggesting that upregulation of Sst mRNA (by 24 hr; Figure 3B) and SST protein in this assay represent immediate early effects of Satb1 misexpression in interneuron precursors.
The separated plasma was collected using a plastic pastette and transferred to clean tubes for immediate storage at −80 °C until assayed using radioimmuno assay with commercially available kits according to the manufacturer's instructions (EURIA-CCK, Euro-Diagnostica, obtained from Immunodiagnostic Systems, Ids Ltd., Tyne and Wear, UK) as described previously.
Together, these assays provide a suite of novel, complementary functional assays with immediate relevance to cancer progression and enabling rapid evaluation of new therapeutic agents, as exemplified below.
