Microsoft word - esk5683.doc

Instruction
This kit is only for scientific research, and shall not be used as a clinical diagnosis of use. This kit allows for the determination of resistin concentrations in Human serum, Urine and other biological fluids. Principle
The kit assay Human Prednisolone, Ps level in the sample，add Human Prednisolone, Ps antibody to microtiter plate wells, after Incubating,add Biotinylated anti Prednisolone, Ps antibody , then Combined Streptavidin-HRP, become complex, after Incubating and washing Completely, Add TMB substrate solution,TMB substrate becomes blue color, reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm. The concentration of Prednisolone, Ps in the samples is then determined by comparing the O.D. of the samples to the Materials provided with kit
Materials provided with the kit
48 determinations
96 determinations
Materials required but not supplied
Precision pipettes and Disposable pipette tips. Important notes
1. The kit takes out from the refrigeration environment should be balanced 15-30 minutes in the room temperature, ELISA plates coated if has not used up after opened, the plate should be stored in Sealed bag. Add Sample with sampler each step, And proofread its accuracy frequently, avoids the experimental error. Please according to use instruction strictly, The test result determination must take the microtiter plate reader as a 4. Use new disposal plastic pipette tips and Closure plate membrane for each standard, in order to avoid cross Do not mix reagents with those from other lots. The substrate evades the light preservation. Specimen requirements
1. Extract as soon as possible after Specimen collection, and according to the relevant literature, and should be experiment as soon as possible after the extraction. If it can't, Specimen can be kept in -20ºC to preserve, Avoid Can’t detect the sample which contain NaN3,because NaN3 inhibits HRP active. Assay procedure
Dilute and add sample: Dilute Original density Standard as follow table: 120μl original density Standard+120μl Standard diluent 120μl 5 Standard +120μl Standard diluent 120μl 4 Standard +120μl Standard diluent 120μl 3 Standard +120μl Standard diluent 120μl 2 Standard +120μl Standard diluent 2. According to testing Sample numbers to define how many wells needed ,Standard and blank suggested repeated holes. Quantity determination in accordance with samples, and do repeated holes as far as possible. sample：1) blank wells: (blank comparison wells don’t add sample , Biotinylated anti –Prednisolone, Ps-antibody and Streptavidin-HRP ,other each step operation is same); 2) Standard wells: add Standard 50μl and Streptavidin-HRP 50μl; 3) testing Sample wells: add sample 40μl,then add anti –Prednisolone, Ps-antibody 10μl , Streptavidin-HRP 50μl. closing plate with Closure plate membrane ,incubate for 60 min at 37ºC. Reagent preparation:30-fold(or 20-fold) wash solution diluted 30-fold(or 20-fold) with distilled water and reserve. Washing: Uncover closure plate membrane, discard Liquid, dry by swing, add washing buffer to every well, still for 30s then drain, repeat 5 times, then pat dry. Chromogenic reaction: add chromogenic agent solution A 50μl and chromogenic agent solution B to each well, evade the light preservation for 15 min at 37ºC Stop the reaction: add stop solution 50μl to each well, stop the reaction(the blue color change to yellow color). Assay: take blank well as zero, read absorbance at 450nm after adding stop solution and within 10min. Step description
Add Standard. sample diluent, Biotinylated and HRP, incubate for 60 min at 37 ºC. Wash 5 times, add Chromogenic agent Solution A and B, incubate for 15 min at 37 ºC. Storage and validity

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