ELISA assay was performed using antigen at 0.5µg/ml and antibody at 1µg/ml. ELISA plates coated on bench overnight in 0.1M Carbonate pH 9.6, 100µl/well, washed once in wash buffer 300µl/well (TBS + 0.1% tween 20) and blocked 2 hours in 1% BSA in D-PBS 300µl/well. Antibodies diluted to working strength in diluent (DPBS + 1% BSA + 0.05% Tween 20 + 0.2% Proclin 950). Added at 100µl/well and incubated 2 hours shaken at ambient temperature. Washed 3 x 300µl/well. Goat anti Mouse IgG-HRP (Biorad103005) diluted 1 in 2500 in diluent, added at 100µl/well and incubated with shaking 1 hour at ambient. Plate washed 6X300µl/well. TMB (KPL Sureblue 5120-0077) added at 100µl/well. Reaction for screening assay stopped by addition of 1M HCl 100µl/well.
Image by Virology Research Services Ltd
Immunofluorescence was carried out in Vero cells. Cells were seeded on coverslips and infected with DENV (serotypes 1, 2, 3 and 4) for 48h at MOI 1. Control coverslips consisted of uninfected Vero cells. After fixation with 4% PFA, samples were stained with Mouse anti-Dengue virus NS1 antibody (AbDENV14NS1-CM436). Antibody was diluted 1:500 and Triton X-100 was used as detergent. Imaging was performed using a Leica SP5 confocal microscope. Immunofluorescence was observed in DENV serotype 4 infected cells using AbDENV14NS1-CM436, as shown above (Virology Research Services Ltd).
MOUSE ANTI-DENGUE VIRUS NS1 SEROTYPE 4 ANTIBODY (CM436)
Mouse anti Dengue virus serotype 4 NS1 monoclonal antibody is specific for the NS1 protein of Dengue virus serotype 4. It does not cross-react with Dengue NS1 from other serotypes, nor with NS1 from Zika virus or other flaviviruses. No cross-reactivity is seen with Chikungunya virus proteins (E1, E2 and C). This mouse anti Dengue virus serotype 4 NS1 antibody is suitable for use in direct ELISA and in sandwich ELISA.
PRODUCT DETAILS – MOUSE ANTI-DENGUE VIRUS NS1 SEROTYPE 4 ANTIBODY (CM436)
- Mouse anti-Dengue virus NS1 serotype 4 monoclonal antibody (clone CM436).
- Greater than 95% purity by SDS-PAGE and buffered in PBS, pH7.4.
BACKGROUND
The NS1 protein is a major non-structural protein expressed by the Dengue Virus. The NS1 monomer is a glycosylated protein of approximately 45kD, which associates with lipids and forms a homodimer inside infected cells. It is necessary for viral replication, and is also secreted into the extracellular space as a hexameric lipoprotein particle, which is involved in immune evasion and pathogenesis by interacting with components from both the innate and adaptive immune systems, as well as other host factors. NS1 is one of the major antigenic markers for viral infection with Dengue.
With more than one-third of the world’s population living in areas at risk of transmission, dengue infection is a leading cause of illness and death in the tropics and subtropics (WHO for Guidelines for Diagnosis, Treatment, Prevention & Control, 2009) As many as 100 million people are infected yearly. Dengue is caused by any one of four related viruses transmitted by mosquitoes. There are no vaccines available to prevent infection with dengue virus.
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