Ferritin, Human, BioAssay(TM) ELISA Kit

Ferritin is a ubiquitous and highly conserved protein which plays a major role in iron homeostasis. It is a holoenzyme shell (~450 kD) consisting of 24 subunits of two types, H(heavy) and L(light), and capable of storing up to 4,500 atoms of ferric iron. Depending on the tissue type and physiologic status of the cell, the ratio of H to L subunits in ferritin can vary widely. It can be viewed not only as part of a group of iron regulatory proteins that include transferrin and the transferrin receptor, but also as a member of the protein family that orchestrates the cellular defense against stress and inflammation. Ferritin is found in the liver, spleen, kidney and heart. Only a small amount is found in the blood. The blood level of ferritin serves as an indicator of the amount of iron stored in the body.\n\nIntended Use:\nHuman Ferritin ELISA kit is to be used for the in vitro quantitative determination of human Ferritin in human serum, human plasma, cell lysate and buffered solution. The assay will recognize native Ferritin. This kit has been configured for research use only and is not to be used in diagnostic procedures.\n\nSensitivity:\nThe minimal detectable dose of human Ferritin was calculated to be 0.0104ng/ml, by subtracting two standard deviations from the mean of 12 zero standard replicates (ELISA buffer, S0) and intersecting this value with the standard curve obtained in the same calculation.\n\nSpecificity:\nThe following substances were tested and found to have no cross-reactivity: human serum albumin, human alpha fetoprotein(AFP), transferrin, plasminogen, human Vitamin D binding protein(VDBP), Hemoglobin, Human IgG.\n\nTest Principle:\nThe design of this assay is based on a sandwich Enzyme-Linked Immunosorbent Assay (ELISA). The microtiter plate provided in this kit has been pre-coated with a polyoclonal antibody specific to human Ferritin. Samples are pippetted into these wells. Nonbound Ferritin and other components of the sample should be removed by washing, then biotin-conjugated polyclonal antibody specific to Ferritin added. In order to quantitatively determine the amount of Ferritin present in the sample, Avidin conjugated to Horseradish Peroxidase (HRP) should be added to each microplate well. The final step, a TMB-substrate solution added to each well. Finally, a sulfuric acid solution is added and the resulting yellow colored product is measured at 450nm. Since the increases in absorbency is directly proportional to the amount of captured Ferritin.\n\nKit Components:\nMicroplate: 1x96 wells\nIncubation buffer: 1x30ml\nWashing buffer, (10x): 1x100ml\nStandard protein: 1 glass vial lyophilized\nStandard/sample dilution buffer: 1x25ml\nSecond antibody: 1 glass vial lyophilized\nAV-HRP, (100x): 1x150ul\nSecondary antibody/AV-HRP dilution buffer: 1x25ml\nSubstrate (TMB): 1x20ml\nStop solution: 1x20ml\n\nStorage and Stability:\nStore components at 4 degrees C. Stable for at least 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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SPECIFICATIONS

Catalog Number

F4015-18Q

Size

96Tests

References

1. Theil, E.C. (2003) J. Nutr. 133:1549-1553. 2. Torti, F.M. and Torti, S.V. (2002) Blood, 99:3505-3516. 3. Arosio, P. and Levi, S. (2002) Free Radic. Biol. Med. 33(4):457-463