Description
FUT-175 (Futhan, or Nafamostat Mesilate, chemical name: 6-amidino-2-naphthyl p-guanidinobenzoate dimethanesulfonate) is a synthetic, broad-specificity protease inhibitor and an inhibitor of the classical and alternate pathways of complement activation. The molecular weight of Futhan is 539.58 Da.
Preparation And Storage
Protect from light and moisture. After reconstitution with dH2O, Futhan is stable for 4 weeks if stored at 4°C and several months if stored at -80°C.
Recommended Assay Procedures
Complement activation in vivo may initiate, contribute to, or exacerbate the inflammatory reactions seen in gram-negative bacterial sepsis, trauma, ARDS, ischemic heart disease, post-dialysis syndrome and several autoimmune diseases including rheumatoid arthritis, lupus erythematosus and acute glomerulonephritis. Although chelation of bivalent cations by EDTA prevents the activation of a number of plasma proteases, including the coagulation and complement pathways, it has been reported that cleavage of certain complement components still occurs. This makes in vitro measurements of the in vivo-generated cleavage products, such as C3a, C4a, or C5a, less accurate. Addition of FUT-175 (Futhan) to plasma samples at the time of sample collection provides additional protection from ex-vivo activation, and therefore ensures more accurate measurements that reflect the circulating levels of complement activation products.
Sample collection for anaphylatoxin measurements: Prior to use, reconstitute FUT-175 (Futhan) with 1 ml dH2O to get a 100x stock solution. Avoid using PBS or other phosphate-containing buffers as it might reduce the solubility of FUT-175. Add 10 µl of FUT-175 (Futhan) stock per ml of freshly drawn EDTA blood (i.e. 100 µl FUT-175 (Futhan) stock/10 ml EDTA blood). Keep blood sample on ice, then spin and collect plasma. Proceed to C3a-desArg, C4a-desArg or C5a-desArg measurements using our OptEIA kits (Cat No. 550499, 550947, or 550500), or our mouse complement antibodies for ELISA (558250, 558251, 558618, 558027, 558028, 622597), or store samples at 4°C or frozen at -80°C for later evaluation. Alternatively, FUT-175 (Futhan) can act as a sample stabilizer when it is added at 10 µl/ml to plasma or serum samples. Our studies show that ex vivo generation of C4a-desArg in EDTA plasma samples is blocked up to 24 hours at 4°C or at RT, and up to 5 hours at 37°C, in the presence of the recommended concentration of FUT-175 (Futhan) (i.e. 50 µg/ml).
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Development References (7)
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Hecke F, Schmidt U, Kola A, Bautsch W, Klos A, Kohl J. Circulating complement proteins in multiple trauma patients--correlation with injury severity, development of sepsis, and outcome. Crit Care Med. 1997; 25(12):2015-2024. (Biology). View Reference
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Ikari N, Sakai Y, Hitomi Y, Fujii S. New synthetic inhibitor to the alternative complement pathway. Immunology. 1983; 49(4):685-691. (Biology). View Reference
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Inagi R, Miyata T, Maeda K, Sugiyama S, Miyama A, Nakashima I. FUT-175 as a potent inhibitor of C5/C3 convertase activity for production of C5a and C3a. Immunol Lett. 1991; 27(1):49-52. (Biology). View Reference
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Issekutz AC, Roland DM, Patrick RA. The effect of FUT-175 (Nafamstat Mesilate) on C3a, C4a and C5a generation in vitro and inflammatory reactions in vivo. Int J Immunopharmacol. 1990; 12(1):1-9. (Biology). View Reference
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Pfeifer PH, Kawahara MS, Hugli TE. Possible mechanism for in vitro complement activation in blood and plasma samples: futhan/EDTA controls in vitro complement activation. Clin Chem. 1999; 45(8):1190-1199. (Biology). View Reference
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Stove S, Welte T, Wagner TO, et al. Circulating complement proteins in patients with sepsis or systemic inflammatory response syndrome. Clin Diagn Lab Immunol. 1996; 3(2):175-183. (Biology). View Reference
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Watkins J, Wild G, Smith S. Nafamostat to stabilise plasma samples taken for complement measurements. Lancet. 1989; 1(8643):896-897. (Biology). View Reference
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
