The Human Resistin ELISA Quantification Kit quantifies the amount of resistin in a sample based on a two-site sandwich ELISA method for the determination of resistin. Antibodies against the opposite sex have been pre-coated on 96-well plates. The target protein in the standard and sample binds to the immobilized antibody, and after the unbound substrate is removed, the biotinylated detection antibody of the anisotropy of the target protein is added, and after washing, a brick-rich streptavidin-HRP conjugate is added to the reaction wells. Wash to remove all unbound streptavidin-HRP reagent and add HRP substrate solution to produce a colored product with a shade of color proportional to the content of the protein of interest.
Human ELISA kit procedure.
1.Bring the kit to room temperature for 30 minutes before use.
2.Take the required amount of microplate labels, set up 1 well of blank control, 2 wells of negative and positive control, seal the unused plate as soon as possible, and store 2 8.
3.Blank control wells with 100 l of sample diluent;100 l of negative and positive control were added to the negative and positive control wells, respectivelyAdd 100 l of the diluted sample per well.
4.Mix well and let the 37 reaction for 30 min.
5.Remove the liquid in the wells, fill each well with washing liquid, let stand for 30 seconds and then discard, repeat the washing 5 times, and pat dry.
6.Add 100 l of enzyme marker per well (except blank wells). Set 37 for 30 min.
7.Polyester, same as step 5.
8.50 L of substrate solution A and 50 l of substrate solution B were added to each well, mixed well, and 37 were protected from light for 10 minutes.
9.50 L of stop solution was added to each well, mixed well, zeroed with a blank well, and the absorbance value (A value) of each well was measured at 450 nm (630 nm can be used as a reference wavelength).
Precautions in use:
1.The reagents in the kit should not be mixed with reagents from other lot numbers or reagents that are out of expiration date.
2.Reagents should be stored as labeled on the bottle and equilibrated at room temperature for 30 minutes before use.
3.Use disposable tips to avoid cross-contamination.
4.The slats that were not used in the experiment were immediately put back into the package, sealed tightly, and preserved.4
5.Thorough mixing is especially important for reaction results, and it is recommended to use a low-speed frequency shaker or hand shaking every 10 minutes.
6.For samples and standards, it is recommended to do two- or three-well testing.