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Ferritin CLIA Kit

FE Reactivity: Human Chemiluminescent Sandwich ELISA
Catalog No. ABIN504790
  • Target See all Ferritin (FE) CLIA Kits
    Ferritin (FE)
    Reactivity
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    • 2
    • 2
    • 1
    • 1
    • 1
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    Human
    Detection Method
    Chemiluminescent
    Method Type
    Sandwich ELISA
    Application
    ELISA
    Purpose
    Immunoenzymometric sequential assay: The essential reagents required for an immunoenzymometric assay include high affinity and specificity antibodies (enzyme and immobilized), with different and distinct epitope recognition, in excess, and native antigen. In this procedure, the immobilization takes place during the assay at the surface of a microplate well through the interaction of streptavidin coated on the well and exogenously added biotinylated monoclonal anti- ferritin antibody. Upon mixing monoclonal biotinylated antibody, and a serum containing the native antigen, reaction results between the native antigen and the antibody, forming an Antibody-Antigen complex. Simultaneously the biotin attached to the antibody binds to the streptavidin coated on the microwells resulting in immobilization of the complex.
    Analytical Method
    Quantitative
    Characteristics
    The Quantitative Determination of Circulating Ferritin Concentrations in Human Serum by a Microplate Chemiluminescence immunoassay (CLIA)
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  • Application Notes
    All products that contain human serum have been found to be non-reactive for Hepatitis B Surface Antigen, HIV 1&2 and HCV Antibodies by FDA required tests. Since no known test can offer complete assurance that infectious agents are absent, all human serum products should be handled as potentially hazardous and capable of transmitting disease. Good laboratory procedures for handling blood products can be found in the Center for Disease Control / National Institute of Health, "Biosafety in Microbiological and Biomedical Laboratories," 2nd Edition, 1988, HHS Publication No. (CDC) 88-8395.
    Sample Volume
    25 μL
    Plate
    Pre-coated
    Protocol

    Specimien Collection and Preparation:

    The specimens shall be blood, serum in type and the usual precautions in the collection of venipuncture samples should be observed. For accurate comparison to established normal values, a fasting morning serum sample should be obtained. The blood should be collected in a plain redtop venipuncture tube without additives or anti-coagulants. Allow the blood to clot. Centrifuge the specimen to separate the serum from the cells. Samples may be refrigerated at 2_x001E_8(C for a maximum period of five (5) days. If the specimen(s) cannot be assayed within this time, the sample(s) may be stored at temperatures of _x001E_20(C for up to 30 days. Avoid repetitive freezing and thawing. When assayed in duplicate, 0.050ml of the specimen is required.

    Reagent Preparation:

    1. Wash Buffer Dilute contents of Wash Concentrate to 1000ml with distilled or deionized water in a suitable storage container. Store diluted buffer at room temperature 20-27(C. 2. Working Signal Reagent Solution - Store at 2 - 8(C. Determine the amount of reagent needed and prepare by mixing equal portions of Signal Reagent A and Signal Reagent B in a clean container. For example, add 1 ml of A and 1ml of B per two (2) eight well strips (A slight excess of solution is made). Discard the unused portion if not used within 36 hours after mixing. If complete utilization of the reagents is anticipated, within the above time constraint, pour the contents of Signal Reagent B into Signal Reagent A and label accordingly.

    Test Procedure:

    Before proceeding with the assay, bring all reagents, serum references and controls to room temperature (20 - 27( C). 1. Format the microplates wells for each serum reference, control and patient specimen to be assayed in duplicate. Replace any unused microwell strips back into the aluminum bag, seal and store at 2-8(C. 2. Pipette 0.025 ml (25l) of the appropriate serum reference, control or specimen into the assigned well. 3. Add 0.100 ml (100l) of the Ferritin Biotin Reagent to each well. It is very important to dispense all reagents close to the bottom of the coated well. 4. Swirl the microplate gently for 20-30 seconds to mix and cover. 5. Incubate 30 minutes at room temperature. 6. Discard the contents of the microplate by decantation or aspiration. If decanting, tap and blot the plate dry with absorbent paper. 7. Add 350l of wash buffer (see Reagent Preparation Section), decant (tap and blot) or aspirate. Repeat four (4) additional times for a total of five (5) washes. An automatic or manual plate washer can be used. Follow the manufacturers instruction for proper usage. If a squeeze bottle is employed, fill each well by depressing the container (avoiding air bubbles) to dispense the wash. Decant the wash and repeat four (4) additional times. 8. Add 0.100ml of Ferritin Tracer Reagent. It is very important to dispense all reagents close to the bottom of the coated well. 9. Incubate 30 minutes at room temperature. 10. Discard the contents of the microplate by decantation or aspiration. If decanting, tap and blot the plate dry with absorbent paper. 11. Add 350l of wash buffer (see Reagent Preparation Section), decant (tap and blot) or aspirate. Repeat four (4) additional times for a total of five (5) washes. An automatic or manual plate washer can be used. Follow the manufacturers instruction for proper usage. If a squeeze bottle is employed, fill each well by depressing the container (avoiding air bubbles) to dispense the wash. Decant the wash and repeat four (4) additional times. 12. Add 0.100 ml (100l) of working signal reagent to all wells (see Reagent Preparation Section). ). Always add reagents in the same order to minimize reaction time differences between wells. 12. Incubate for five (5) minutes at room temperature in the dark. 13. Read the relative light units in each well for 0.2 1.0 seconds. The results should be read within thirty (30) minutes of adding the substrate solution.
    Restrictions
    For Research Use only
  • Target See all Ferritin (FE) CLIA Kits
    Ferritin (FE)
    Alternative Name
    Ferritin (FE Products)
    Synonyms
    1HCH CLIA Kit, BEST:LD36673 CLIA Kit, CG2216 CLIA Kit, Dmel\\CG2216 CLIA Kit, FER1 CLIA Kit, FER1HCH CLIA Kit, Fer-L CLIA Kit, Fer1 CLIA Kit, Fer1HC CLIA Kit, FerHCH CLIA Kit, HCH CLIA Kit, T21 CLIA Kit, T9 CLIA Kit, fer1 CLIA Kit, fer1hch CLIA Kit, l(3)00451 CLIA Kit, l(3)j10B4 CLIA Kit, SFerH-1 CLIA Kit, SOF-35 CLIA Kit, SOF-5L CLIA Kit, NV13011 CLIA Kit, ACP1 CLIA Kit, F8A5.24 CLIA Kit, F8A5_24 CLIA Kit, Ferritin 1 heavy chain homologue CLIA Kit, ferritin light chain CLIA Kit, ferritin CLIA Kit, chromosome 26 open reading frame, human C10orf131 CLIA Kit, hyaluronan and proteoglycan link protein 2 CLIA Kit, NAD(P)-linked oxidoreductase superfamily protein CLIA Kit, Fer1HCH CLIA Kit, H-1 CLIA Kit, YPDSF_1340 CLIA Kit, ftnA CLIA Kit, Ft CLIA Kit, C26H10orf131 CLIA Kit, HAPLN2 CLIA Kit, AT1G60730 CLIA Kit
    Background
    Ferritin, in circulation, as measured in serum levels is a satisfactory index of bodys 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 bodys 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, leukemias, 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. In this method, ferritin calibrator, patient specimen or control is first added to a streptavidin coated well. Biotinylated monoclonal antibody (specific for ferritin) is added and the reactants mixed. Reaction results between the biotinylated ferritin antibody and native ferritin to form an immune complex that is deposited on the streptavidin coated well. The excess serum proteins are washed away via a wash step. Another ferritin specific antibody, labeled with an enzyme, is added to the wells. The enzyme labeled antibody binds to the ferritin already immobilized on the well. Excess enzyme is washed off via a wash step. A light signal is generated by the addition of a substrate. The intensity of the light generation is directly proportional to the concentration of the ferritin in the sample. The employment of several serum references of known ferritin levels permits the construction of a dose response curve of activity and concentration. From comparison to the dose response curve, an unknown specimen's activity can be correlated with ferritin concentration.
    Pathways
    Transition Metal Ion Homeostasis
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