Darbepoetin ELISA Assay Kit


The Darbepoetin ELISA Assay Kit is for the quantitative determination of Darbepoetin (Aranesp) in serum, plasma and cell culture samples. The Darbepoetin ELISA Assay Kit is intended for research use only and not for use in diagnostic or clinical procedures.

Darbepoetin ELISA Assay Kit

Darbepoetin ELISA Assay Kit

Size: 12×8 wells
Sensitivity: 100pg/ml
Standard Range: 100pg/ml – 20ng/ml
Incubation Time: 4 hours
Sample Type: Serum, plasma
Sample Size: 100µl
Alternative Names: ARANESP

Assay Principle

Erythropoietin (EPO) is a heavily glycosylated protein with a molecular weight of about 30,000 – 34,000 Daltons. Human EPO is a polypeptide consisting of 165 amino acids, containing one O-linked and three Nlinked carbohydrate chains. The recombinant EPO is a good substitute for the native protein for use in an immunoassay. Darbepoetin is a re-engineered form of erythropoietin, contains 5 N-linked oligosaccharide chains and has a molecular weight of 37,100 Daltons and a carbohydrate composition of 51%.It has a 3-fold longer serum half-life compared to epoetin alpha and epoetin beta.

Sample Preparation and Storage:
Blood is taken by venipuncture. Serum is separated after clotting by centrifugation. Plasma can be used, too. Lipaemic, hemolytic or contaminated samples should not be run. Repeated freezing and thawing should be avoided. If samples are to be used for several assays, initially aliquot samples and keep at – 20 °C.
Preparation Before Use:
Allow samples to reach room temperature prior to assay. Take care to agitate patient samples gently in order to ensure homogeneity.
Serum/Plasma/ Dilution: Add 2.0µl of Serum or Plasma to 398.0µl of Assay diluent.
Limit Of Detection: It is defined as the lowest detectable concentration corresponding to a signal of Mean of ‘0’ standard plus 2* SD.
10 replicates of ‘0’ standards were evaluated and the LOD was found to be less than 100 pg/ml.

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Additional Information

Assay Principle

The method employs the quantitative enzyme immunoassay technique. Standards or samples are pipetted into microwells pre-coated with Anti-Darbepoetin antibody and Darbepoetin present in the sample and standards are bound by Anti- Darbepoetinantibody . In the second step, Detection antidody is added and incubated. In the third step, a HRP conjugate is pipetted and incubated. Free HRP conjugate will be removed by washing. Addition of TMB substrate will develop blue color and intensity of blue colour in wells is proportional to theconcentration of Darbepoetin present in standard or sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.

Package Inserts

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