This kit is designed around a hexon-specific antibody, which is used to label infected cells. Hexon protein is encoded by the adenoviral genome and is an essential component of the adenoviral capsid required for adenoviral replication, but its expression depends on the E1 gene product. Thus, E1 trans-complementing cell types such as HEK 293 can be used to measure infectious activity because only infected cells will produce the hexon protein. The Rapid Titer assay takes 1–2 hours to set up and another 3 hours (two days later) to label, stain, and count infected cells. Each stained cell corresponds to a single infectious unit.
Adeno-X qPCR Titration Kit
The Adeno-X qPCR Titration Kit provides a fast, simple, and accurate method for titrating adenoviral stocks from all Ad5-based adenoviral vectors, including recombinant adenovirus created with our Adeno-X Expression Systems. The protocol combines qPCR and TB Green intercalating dye chemistry, allowing you to determine the viral genome copy number in adenoviral preparations (i.e., crude lysates or purified stocks) from a calibrated DNA standard curve. The kit delivers results in just 4 hours, and because qPCR titration is so fast, target cells can be infected with accurately titrated virus on the same day the virus is harvested.
The GoStix test, which resembles a pregnancy test but instead detects adenoviral hexon proteins, requires just 20 µl of adenoviral supernatant to determine if infectious virus was produced posttransfection. Adeno-X GoStix are also ideal tools for determing the best time to harvest your virus following amplification, since the amount of adenovirus detected in the supernatant correlates strongly with adenovirus production inside the cells. More information on Adeno-X GoStix.