immunoassay control group 10  (R&D Systems)

Journal: Journal of Virology

Article Title: Rabies Virus Stimulates Nitric Oxide Production and CXC Chemokine Ligand 10 Expression in Macrophages through Activation of Extracellular Signal-Regulated Kinases 1 and 2

doi: 10.1128/jvi.78.17.9376-9388.2004

Figure Lengend Snippet: FIG. 4. RV stimulates gene expression and protein production of CXCL10 in macrophages. (A) Time course of CXCL10 transcription in macrophages. Cells were incubated with () or without () infectious and UV-inactivated (Inact.) CVS virions for 2 h. After additional incubation for the indicated times, CXCL10 mRNAs from each sample were amplified by using RT-PCR as described in the text. The data are from one of three independent experiments with similar results. (B) Measurement of the relative amounts of CXCL10 mRNA in macrophages upon RV stimulation. RAW264 cells were stimulated with RV (10 FFU/cell), washed, and further incubated. Immediately thereafter (0 h) and at the time points indicated, CXCL10 mRNAs were detected by using RT-PCR, and the relative amounts of CXCL10 mRNA were determined by image analysis of the band intensity of each PCR product as described in the text. The results are shown as n-fold increase in the expression of CXCL10 mRNA with reference to the levels of CXCL10 mRNA at 0 h after incubation. Mean values and standard deviations from the results of three independent experiments are shown. (C) Protein contents of CXCL10 in culture fluids from RV-stimulated macrophages. Cells were stimulated with RV inocula (10 FFU/ml) for 2 h or unstimulated (Mock) and then incubated for the times indicated. After incubation, the culture supernatants were removed, and the CXCL10 protein content was measured by enzyme-linked immunosorbent assay. The data are averages of three independent experiments, and the error bars represent standard deviations.

Article Snippet: After incubation for the appropriate times, the culture fluids were separated, and the levels of CXCL10 protein content were determined by using an enzyme-linked immunosorbent assay kit (Quantikine mouse IP-10 immunoassay; R&D Systems Inc., Minneapolis, Minn.) according to the manufacturer’s instructions.

Techniques: Gene Expression, Incubation, Reverse Transcription Polymerase Chain Reaction, Expressing, Enzyme-linked Immunosorbent Assay