This kit is used to assay the Erythropoietin(EPO)in the sample of Human’s serum, blood plasma, and other related tissue Liquid.
The kit uses a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) to assay the level of Human Erythropoietin(EPO)in samples. Add Erythropoietin(EPO) to monoclonal antibody Enzyme well which is pre-coated with Human Erythropoietin(EPO)monoclonal antibody, incubation; then, add Erythropoietin(EPO)antibodies labeled with biotin, and combined with Streptavidin-HRP to form immune complex; then carry out incubation and washing again to remove the uncombined enzyme. Then add Chromogen Solution A, B, the color of the liquid changes into the blue, And at the effect of acid, the color finally becomes yellow. The chroma of color and the concenthumanion of the Human Substance Erythropoietin(EPO)of sample were positively correlated.
Materials supplied in the Test Kit
1 Standard(800mIU/ml) 0.5ml 7 Chromogen Solution A 6ml
2 Standard diluent 3ml 8 Chromogen Solution B 6ml
3 Microelisa Stripplate 12w×8s 9 Stop Solution 6ml
4 Str- HRP-Conjugate Reagent 6ml 10 Instruction 1
5 30×wash solution 20ml 11 Closure plate membrane 2
6 Biotin-EPO Ab 1ml 12 Sealed bags 1
Materials required but not supplied
1. 37 ℃ incubator 2. Standard Enzyme reader
3. Precision pipettes and Disposable pipette tips 4. Distilled water
5. Disposable tubes for sample dilution 6. Absorbent paper
1. Beening taken out from the 2-8℃ environment, the kit should be balanced 30 minutes in the ambient temperature then use. If the Coated plates of Enzyme haven’t been used up after opened, the remaining plates should be stored in Sealed bag.
2. For each step, add Sample with sample injector which should be calibrated frequently, in order to avoid unnecessary experimental tolerance.
3. he operation shall be carried out accordance to the instructions strictly. And test results must be based on the readings of the Enzyme reader.
4. In order to avoid cross-contamination, it is forbidden to re-use the suction head and seal plate membrane in your hands.
5. All samples, washing buffer and each kind of reject should according to infective material process.
6. The idle agents shall be put up or covered. Do not use reagent with different batches. And use them before expired date.
7. The substrate B is light-sensitive. Prolonged exposure to light is forbidden.
Manually washing method: shake away the remain liquid in the enzyme plates; place some bibulous papers on the test-bed, and flap the plates on the upside down strongly. Inject at least 0.35ml after-dilution washing solution into the well, and marinate 1~2 minutes. Repeat this process according to your requirements.
Automatic washing method: if there is automatic washing machine, it should only be used in the test when you are quite familiar with its function and performance.
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Human EPO were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Human EPO were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100