The DENV is small structure that can only reproduce inside a host organism. There are 4 related dengue viruses- Degue-1, Dengue-2, Dengue-3, & Dengue4. All of them are present in the same areas of the world. The dengue virus has just about a sphere shaped structure comprised of the viral genome and capsid proteins bordered by an envelope and a shell of antigen. Subsequently infecting, the dengue virus takes over the host cell's system to replicate the viral RNA genome and viral antigens. Once matured, the resynthesized DENVs are discharged and continue to infect additional host cells.
Recombinant Dengue-HRP is genetically designed dengue protein conjugated to Horseradish peroxidase which detects low titer dengue antibodies having a sharp OD value compared to the control with optimal dilutions. The proprietary Dengue-HRP protein sequence is designed for broad reactions to specific antibodies produced from the different serotypes of dengue infection. Potential applications for Dengue-HRP include indirect ELISA for the detection of dengue IgG or MAC ELISA for the detection of dengue IgM.
Phosphate buffered saline, pH 7.4 and 25% glycerol.
Store at 4°C if entire vial will be used within 2-4 weeks. Store, frozen at -20°C for longer periods of time. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA). Avoid multiple freeze-thaw cycles.
Indirect ELISA and MAC-ELISA 1:100-200 diluted Dengue-HRP produced 3 to 6 fold OD value compared to OD from the control. The OD value may vary depending on the concentration of specific IgG or IgM, sample fold dilution, sample loading volume and other variables.
Coat each well with 1 to 3ug/well polyclonal or monoclonal anti-human IgG or IgM. Leave in 4C for overnight. Wash with wash buffer and block with 3% goat serum PBS for 1 hour at 37 C. Wash with wash buffer again. 1:20 dilution of sample with PBS. Add 100ul diluted sample into each well. Leave in 37 oC for 1 hour. Wash with wash buffer again. 1:100 to 200 dilute Den-HRP with PBS and add 100ul into each well. Incubate for 1 hour at 37C. After wash with wash buffer and add 100ul substrate for color reaction.