DRG Hepcidin 25 (bioactive) HS Elisa Kit

PRINCIPLE OF THE TEST
The DRG Hepcidin 25 (bioactive) HS ELISA Kit is a solid phase enzyme-linked immunosorbent assay (ELISA) based on the principle of competitive binding. The microtiter wells are coated with a monoclonal (mouse) antibody directed towards an antigenic site of the Hepcidin-25 molecule. Endogenous Hepcidin-25 of a patient sample competes with a Hepcidin-25-biotin conjugate (Enzyme Conjugate) for binding to the coated antibody. After incubation the microtiter plate is washed to stop the competition reaction. In the following incubation the bound biotin molecules are detected with streptavidin peroxidase (Enzyme Complex). After incubation the plate is washed the second time. After addition of the substrate solution, the intensity of colour developed is inversely proportional to the concentration of Hepcidin-25 in the patient sample.

Storage Conditions
When stored at 2 °C to 8 °C unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date. 
Opened reagents must be stored at 2 °C to 8 °C. Microtiter wells must be stored at 2 °C to 8 °C. Once the foil bag has been opened, care should be taken to close it tightly again. Opened kits retain activity for 8 weeks if stored as described above.

Specimen Collection
Serum: Collect blood by venipuncture (e.g. Sarstedt Monovette for serum), allow to clot, and separate serum by centrifugation at room temperature. Do not centrifuge before complete clotting has occurred. Patients receiving anticoagulant therapy may require increased clotting time.
Plasma: Whole blood should be collected into centrifuge tubes containing anti-coagulant (e.g. Sarstedt Monovette with the appropriate plasma preparation) and centrifuged immediately after collection.

QUALITY CONTROL 
Good laboratory practice requires that controls be run with each calibration curve. A statistically significant number of controls should be assayed to establish mean values and acceptable ranges to assure proper performance. 
It is recommended to use control samples according to state and federal regulations. The use of control samples is advised to assure the day to day validity of results. Use controls at both normal and pathological levels. 
The controls and the corresponding results of the QC-Laboratory are stated in the QC certificate added to the kit. The values and ranges stated on the QC sheet always refer to the current kit lot and should be used for direct comparison of the results. 
Employ appropriate statistical methods for analysing control values and trends. If the results of the assay do not fit to the established acceptable ranges of control materials patient results should be considered invalid. 
In this case, please check the following technical areas: Pipetting and timing devices; photometer, expiration dates of reagents, storage and incubation conditions, aspiration and washing methods. After checking the above mentioned items without finding any error contact your distributor or DRG directly.

WARNINGS AND PRECAUTIONS
This kit is for in vitro diagnostic use only. For professional use only.
All reagents of this test kit which contain human serum or plasma have been tested and confirmed negative for HIV I/II, HBsAg and HCV by FDA approved procedures. All reagents, however, should be treated as potential biohazards in use and for disposal.
Before starting the assay, read the instructions completely and carefully. Use the valid version of instructions for use provided with the kit. Be sure that everything is understood.
The microplate contains snap-off strips. Unused wells must be stored at 2 °C to 8 °C in the sealed foil pouch and used in the frame provided.
Pipetting of samples and reagents must be done as quickly as possible and in the same sequence for each step.