Ferritin ELISA Kit

Sample

Serum

Species Reactivity

Human

Detection Method

sELISA

Intended Use

The Quantitative Determination of Circulating Ferritin Concentrations in Human Serum by a Microplate Enzyme Immunoassay, Colorimetric

Contents of Kit

1.Ferritin Calibrators – 1ml / vial
Six (6) vials of Ferritin calibrators at levels of 0(A), 10(B), 50(C), 150(D), 400(E) and 800(F) ng/ml. Store at 2-8°C. A preservative has been added.
Note: The calibrators, human serum based, were calibrated using a reference preparation, which was assayed against the WHO 3^rd IS 94/572
2. Ferritin Biotin Reagent – 13ml/vial
One (1) vial containing biotinylated monoclonal mouse IgG in buffer, dye, and preservative. Store at 2-8°C.
3. Ferritin Enzyme Reagent – 13 ml/vial
One (1) vial containing Horseradish Peroxidase (HRP) labeled anti-ferritin IgG in buffer, dye and preservatives. Store at 2- 8°C.
4. Streptavidin Coated Plate – 96 wells
One 96-well microplate coated with streptavidin and packaged in an aluminum bag with a drying agent. Store at 2-8°C.
5. Wash Solution Concentrate – 20 ml/vial
One (1) vial containing a surfactant in buffered saline. A preservative has been added. Store at 2-8°C.
6. Substrate A – 7ml/vial
One (1) vial containing tetramethylbenzidine (TMB) in buffer. Store at 2-8°C.
7. Substrate B – 7ml/vial
One (1) vial containing hydrogen peroxide (H₂O₂ ) in buffer. Store at 2-8°C.
8. Stop Solution – 8m/vial
One (1) vial containing a strong acid (1N HCl). Store at 2-8°C.
9. Product Instructions.
Note 1: Do not use reagents beyond the kit expiration date.
Note 2: Avoid extended exposure to heat and light. Opened reagents are stable for sixty (60) days when stored at 2-8°C. Kit and component stability are identified on the label.
Note 3: Above reagents are for a single 96-well microplate. For other kit configurations, please refer to the table at the end of the instructions.

Performance Characteristics

Precision
The within and between assay precisions of the ferritin ELISA test system were determined by analyses on three different levels of control sera. The number (N), mean value (X), standard deviation (σ) and coefficient of variation (C.V.) for each of these control sera are presented in Table 2 and Table 3.

Specificity
The cross-reactivity of the ferritin ELISA test system to selected substances was evaluated by adding the interfering substance to a serum matrix at various concentrations. The crossreactivity was calculated by deriving a ratio between dose of interfering substance to dose of Ferritin needed to produce the same absorbance.

Sensitivity
The minimum detectable dose (Sensitivity) is defined as the apparent concentration 2 σ above the absorbance for zero calibrator. 2 σ of the mean absorbance for twenty replicates for zero calibrator for the ferritin ELISA test system gave a sensitivity of 0.17 ng/ml.
High Dose Effect
Since the assay is sequential in design, high concentrations of ferritin do not show the hook effect. Samples with concentrations over 50,000 ng/ml demonstrated extremely high levels of absorbance.

General Description

Ferritin, in circulation, as measured in serum levels is a satisfactory index of body's iron storage. The iron storage is directly measured by quantitative phlebotomy, iron absorption studies, liver biopsies and microscopic examinations of bone marrow aspirates. Iron deficiency (Anemia) and iron overload (Hemochromatosis) are conditions associated with body's iron storage or lack thereof. Measurements of total iron binding capacity (TIBC) have widely been used as aids in the determination of these conditions. However, an assay of serum Ferritin is simply more sensitive and reliable means of demonstration these disorders.
Ferritin is present in blood in very low concentrations. Normally, approximately 1% of plasma iron is contained in Ferritin. The plasma ferritin, is in equilibrium with body stores, and variations of iron storage. The plasma concentrations of ferritin decline very early in anemic conditions like development of iron deficiency, long before the changes are observed in the blood hemoglobin concentration, size of the erythrocytes and TIBC. Thus measurements of serum ferritin can serve as an early indicator of iron deficiency that is uncomplicated by other concurrent conditions. At the same time a large number of chronic conditions can result in elevated levels of serum ferritin. These include chronic infections, chronic inflammatory diseases such as rheumatoid arthritis, heart disease and some other malignancies, especially lymphomas, leukemia, breast cancer and neuroblastoma. In patients who have these chronic disorders together with iron deficiency, serum ferritin levels are often normal. An increase in circulating ferritin is observed in patients with viral hepatitis or after a toxic liver injury as a release of ferritin from the injured liver cells. Elevated serum ferritin levels are found in patients with hemochromatosis and hemosiderosis.
Circulating ferritin levels have been used by clinicians, as an aid, in the diagnosis of several other disorders. It has proved as a valuable tool in differential diagnosis of anemia due to iron deficiency and anemias due to other disorders and, in exposing the depletion of iron reserves long before the onset of anemia. Serial determinations have been used to monitor, non-invasively, the erosion of iron storage during pregnancy and in patients undergoing dialysis. Serum ferritin is routinely used as a screen for iron deficiency for a variety of populations like blood donors and people who are receiving regular blood transfusions or iron replacement therapy.