Elisa Test Kit- RAT, MOUSE, Pack Size 96 T, Temperature Storage 2-8 C

Intended usethis elisa kit applies to the in vitro measurement of quantities for rat ins concentrations in cell culture supernatant and tissue homogenate. Test principle this elisa kit uses the sandwich, Elisa principle. The micro elisa plate provided in this kit has been pre, Layered in an antibody specific to rat ins. Samples or standards are incorporated into the micro elisa plate wells and alongside the specific antibody. Then a biotinylated detection antibody specific for rat ins and avidin, Horseradish peroxidase hrp conjugate are added successively to each micro plate properly nurtured and developed free components are washed away. The substrate solution is included in each well. Only those wells that contain rat ins, Biotinylated detection antibody and avidin, Hrp conjugate will appear blue in color. The enzyme, Substrate reaction is terminated through the incorporation of stop solution together with the tint turns yellow. The optical density od is measured spectrophotometrically at a wavelength of 450 2 nm. The od value corresponds to the level of rat ins. You can calculate the concentration of rat ins in the samples by comparing the od of the samples to the standard curve. Sample collectiontissue homogenates It is advised to get detailed references from the literature before analyzing different tissue types. For hemolyzed blood, General information may affect the results, So the tissues should be minced into small pieces and rinsed in ice, Cold pbs 0. 01m, Ph=7. 4 to remove excess blood thoroughly. Tissue pieces should be weighed and then homogenized in pbs tissue weight g pbs ml volume=1 9 with a glass homogenizer on ice. To further break down you can, The cells sonicate the suspension with an ultrasonic cell disrupter or subject it to freeze, Thaw cycles. The homogenates are then centrifuged for 5, 10 min at 5000 g at 2, 8??? to get the supernatant. Cell culture supernatant centrifuge samples for 20 min at 1000 g at 2, 8???. Collect the supernatant to execute the assaynote for kit1 Intended for investigative purposes only. Not suitable for deployment in diagnostic procedures. 2 please wear lab coats, Eye protection and latex gloves for protection. Please perform the experiment following the national security protocols of especially when, Biological laboratories detecting blood samples or other bodily fluids. 3 a freshly opened elisa plate may appear which is, A water, Like substance normal and is unable to have any impact on the experimental results. Return the unused wells to the foil pouch and store as stated by the conditions suggested in the above table. 4 do not reuse the reconstituted standard, Biotinylated detection ab concentrated hrp, Working solution conjugate working solution. The unspent undiluted concentrated biotinylated detection ab 100 and other stock solutions should be stored according to the storage conditions in the above table. 5 the microplate reader should be designed to be installed with a filter that can detect the wave length at 450 10 nm. The optical density should be within 0, 3. 5. Follow the instructions of the microplate reader for set, Up and preheat it for 15 min before od measurement. 6 do not mix or substitute reagents with those from other lots or sources. 7 change pipette tips in between adding of each standard level, Between sample adding and between reagent adding. Also, Use separate reservoirs for each reagent. 8 the kit should not be used beyond the expiration date on the kit label. Dilution method please predict the concentration array of the sample in advance. If your test sample needs dilution, Please refer to the dilution method as follows for 100 fold dilution one, Step dilution. Add 5 l sample to 495 l sample diluent to yield 100 fold dilution. For 1000 fold dilution