Factor XII (Hageman Factor) BioAssay(TM) ELISA Kit (Antibodies only)

Factor XII (F.XII, also called Hageman Factor) is a glycoprotein that circulates at levels of 40ug/ml in plasma. Activation of F.XII occurs as a consequence of binding to a negatively charged surface or from the proteolysis by kallikrein to produce alphaXIIa which in turn can\nactivate F.XI to F.XIa. alpha-XIIa can be further cleaved by kallikrein to form alphaXIIa which can feed back to activate more prekallikrein to kallikrein and can also activate F.VII and the classical complement pathway. The natural inhibitors of F.XIIa in plasma are, in order of importance, C1-Inhibitor, alpha 2 Antiplasmin and alpha 2 Macroglobulin.\n\nF.XII in plasma and other fluids can be measured by sandwich-style immunoassay. The F.XII is captured onto a microtiter plate coated with affinity purified goat anti-F.XII. The plate is washed to remove unbound protein and the the captured F.XII is then detected with the addition of goat anti-F.XII IgG conjugated to HRP. The plate is again washed and bound antibody conjugate activity is expressed by incubation with substrate, ortho-phenylene diamine (OPD) for a limited time. The color generated is proportional to the concentration of F.XII initially present in the sample.\n\nMaterials Supplied:\nAntibodies for 5x96 well plates\n\nKit Components:\n\nF0019-08A: Capture Antibody 1x500ul Purified by immunoaffinity chromatography. Supplied a liquid in 50% glycerol. For coating ELISA plates.\n\nF0019-08B: Detecting Antibody (HRP) 1x500ul Purified by immunoaffinity chromatography. Supplied a liquid in 50% glycerol. For detection of captured F.XII.\n\nStorage and Stability:\nFor long-term storage, aliquot and store at -20 degrees C. Aliquots are stable for at least 6 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

F0019-08

Size

1Kit

Applications

ELISA

References

1. Silverberg, M. and Kaplan, A.P., Methods in Enzymology 163: 69-80 (1988). 2. Harlow, E., and Lane, D., Detecting and Quantitating Antigens using the Two-Antibody Sandwich Assay, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, p 580 (1988). 3. De La Cadena, R., et al., in Hemostasis and Thrombosis, 3rd Edition, eds. Colman, R.W., Hirsh, J., Marder, V.J., and Salzman, E.W., p 219-240, J.B. Lippincott Co., Philadelphia (1994). 4. Tankersly, D.L., et al., Thrombosis Research 25: 307-317 (1982). 5. Pixley, R.A., et al., JBC 260: 1723-1729 (1985). 6. Nix, B. and Wild, D., in Immunoassays, A Practical Approach, ed. Gosling, J.P., p 239-261, Oxford University Press (2000). 7. NCCLS. Evaluation of the Linearity of Quantitative Analytical Methods; Proposed Guideline - Second Edition. NCCLS Document EP6-P2 (ISBN 1-56238-446-5, NCCLS, Wayne, Pennsylvania USA (2001). 8. FDA Guidance for Industry. Bioanalytical Method Validation; May 2001, www.fda.gov/cder/guidance/index.htm