Rat Creatine Kinase MB Isoenzyme CK-MB ELISA RUO Test Kit

Rat Creatine Kinase MB isoenzyme;CK-MB ELISA Kit

• For the quantitative in vitro determination of Rat Creatine Kinase MB isoenzyme (CK-MB) concentrations in Rat serum, plasma, culture media or any biological fluid.

Principle

  This ELISA kit uses Sandwich-ELISA as the method. The Microelisa stripplate provided in this kit has been pre-coated with an antibody specific to Creatine Kinase MB isoenzyme (CK-MB) Standards or samples are added to the appropriate Microelisa stripplate wells and combined to the specific antibody. Then a Horseradish Peroxidase (HRP)- conjugated antibody specific for Creatine Kinase MB isoenzyme (CK-MB) is added to each Microelisa stripplate well and incubated. Free components are washed away. The TMB substrate solution is added to each well. Only those wells that contain Creatine Kinase MB isoenzyme (CK-MB) and HRP conjugated Creatine Kinase MB isoenzyme (CK-MB) antibody will appear blue in color and then turn yellow after the addition of the stop solution. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm. The OD value is proportional to the concentration of Creatine Kinase MB isoenzyme (CK-MB) . You can calculate the concentration of Creatine Kinase MB isoenzyme (CK-MB) in the samples by comparing the OD of the samples to the standard curve.

Materials provided with the kit

• Dilution of Standards

Ten wells are set for standards in a Microelisa stripplate. In Well 1 and Well 2, 100μl Standard solution and 50μl Standard Dilution buffer are added and mixed well. In Well 3 and Well 4, 100μl solution from Well 1 and Well 2 are added respectively. Then 50μl Standard Dilution buffer are added and mixed well. 50μl solution is discarded from Well 3 and Well 4. In Well 5 and Well 6, 50μl solution from Well 3 and Well 4 are added respectively. Then 50μl Standard Dilution buffer are added and mixed well. In Well 7 and Well 8, 50μl solution from Well 5 and Well 6 are added respectively. Then 50μl Standard Dilution buffer are added and mixed well. In Well 9 and Well 10, 50μl solution from Well 7 and Well 8 are added respectively. Then 50μl Standard Dilution buffer are added and mixed well. 50μl solution is discarded from Well 9 and Well 10. After dilution, the total volume in all the wells are 50μl and the concentrations are 15ng/ml,10 ng/ml,5ng/ml,2.5ng/mland1.25ng/ml, respectively.

Calculation of Results

  Known concentrations of Rat Creatine Kinase MB isoenzyme (CK-MB) Standard and its corresponding reading OD is plotted on the log scale (x-axis) and the log scale (y-axis) respectively. The concentration of Rat Creatine Kinase MB isoenzyme (CK-MB) in sample is determined by plotting the sample’s O.D. on the Y-axis. The original concentration is calculated by multiplying the dilution factor.

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Rat Creatine Kinase MB isoenzyme (CK-MB) were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Rt Creatine Kinase MB isoenzyme (CK-MB) were tested on 3 different plates, 8 replicates in each plate.

CV(%) = SD/meanX100

Intra-Assay: CV<10%

Inter-Assay: CV<12%

Assay range

0.28 ng/ml - 18 ng/ml

Sensitivity:

0.04 ng/ml