This kit is based on Double antigen-Sandwich ELISA detection method and takes 3h assay time. The microplate provided in this kit has been precoated with antigen. Add standard and properly diluted sample into relevant well respectively. After incubation, wash unbound components. Add HRP-detection antigen, then it binds with HV-Ab bound to precoated antigen. Wash unbound components and add TMB substrate solution. Then, TMB was catalyzed by HRP to produce a blue color product that turned yellow after adding a stop solution. Read the O.D. absorbance at 450nm in a microplate reader. Calculate the concentration of HV-Ab in the sample by plotting standard curve. The concentration of the target substance is proportional to the OD450 value.
Step 1: Add 100ul standard or sample into each well, seal the plate and statically incubate for 90 minutes at 37°C.
Washing: Wash the plate three times and immerse for 1min each time.
Step 2: Add 100ul HRP-antigen working solution, seal the plate and statically incubate for 30 minutes at 37°C.
Washing: Wash the plate five times and immerse for 1min each time.
Step 3: Add 90ul TMB substrate solution, seal the plate and statically incubate for 10-20 minutes at 37°C. (Accurate TMB visualization control is required.)
Step 4: Add 50ul stop solution. Read at 450nm immediately and calculate.
Stability
Perform the stability test for the sealed kit at 37°C and 2-8°C and get relevant data.
ELISA kit(n=5)
37°C for 1 month
2-8°C for 6 months
Average(%)
80
95-100
Notes
Specifically binds with HV-Ab , no obvious cross reaction with other analogues.
Usage Notes
Step 1: Take out the required plate wells, add 50ul Cap/Det Ab into each well, then add 50ul Standard or Sample into individual well. (When adding standard or sample, the disposable tip lightly touches the liquid level. Change the disposable tips for different samples and standards.) Gently tap the plate for 10s to ensure thorough mixing then static incubate for 60 minutes at 37°C.
Washing: Wash the plate twice without immersion.
Step 2: Add 100ul HRP-Streptavidin (orange) into each well, seal the plate and static incubate for 30 minutes at 37°C.
Washing: Wash the plate five times without immersion.
Step 3: Add 90ul TMB substrate solution, seal the plate and static incubate for 10-20 minutes at 37°C. (Accurate TMB visualization control is required.)
Step 4: Add 50ul stop solution. Read at 450nm immediately and calculate.
Antigen
HV
Storage
2-8°C(Sealed), Don't cryopreserve.
Reviews of Rat Hantaan Virus (HV) Antibody ELISA Kit