One antibody molecule can be labeled with multiple biotin molecules,
and one biotin molecule can bind a horseradish peroxidase labeled
avidin, thus bringing a large amount of horseradish peroxidase bound
to the antibody, which has higher sensitivity and signal amplification
effect than the traditional direct horseradish peroxidase labeled
antibody.
Purpose
The kit is to assay Rat 4-Hydroxynonenal (4-HNE) levels in Rat serum, plasma, culture media or any biological fluid.
Principle
The Microelisa stripplate has been pre-coated with an monoclonal antibody or antigen and the detecting antibody or antigen is biotinylated in this kit. Samples and biotinylated polyclonal antibody or antigen are added to the appropriate Microelisa stripplate wells in sequence and washed out with TBS or PBS. Then HRP-Avidin is added to each Microelisa stripplate well. The TMB is used for coloration after the HRP-Avidin has been reacted thoroughly and washed out by TBS or PBS. The TMB will appear blue under peroxidase activity then turn yellow after the addition of the stop solution. The depth of color is positively correlated with the factors to be measured in the sample. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm. The OD value is proportional to the concentration of Rat 4-Hydroxynonenal (4-HNE) . You can calculate the concentration of Rat 4-Hydroxynonenal (4-HNE) in the samples by comparing the OD of the samples to the standard curve.
Assay range
1000pmol/ml-15.6pmol/ml
Intra-Assay: CV≤ 8%
Inter-Assay: CV≤ 12%
Recovery: 70 - 110 percent
Sensitivity