4 chloro 7 nitrobenzofurazan


Summary: A benzofuran derivative used as a protein reagent since the terminal N-NBD-protein conjugate possesses interesting fluorescence and spectral properties. It has also been used as a covalent inhibitor of both beef heart mitochondrial ATPase and bacterial ATPase.

Top Publications

  1. Orriss G, Leslie A, Braig K, Walker J. Bovine F1-ATPase covalently inhibited with 4-chloro-7-nitrobenzofurazan: the structure provides further support for a rotary catalytic mechanism. Structure. 1998;6:831-7 pubmed
    ..NBD-Cl inhibits the enzyme by preventing the modified subunit from adopting a conformation that is essential for catalysis to proceed. ..
  2. Klein C, Pillot T, Chambaz J, Drouet B. Determination of plasma membrane fluidity with a fluorescent analogue of sphingomyelin by FRAP measurement using a standard confocal microscope. Brain Res Brain Res Protoc. 2003;11:46-51 pubmed
    ..As a test experiment, we analysed the lateral diffusion of a fluorescent analogue of sphingomyelin and were able to demonstrate its increase upon cholesterol depletion induced by methyl-beta-cyclodextrin (cdx). ..
  3. Terasaki M, Loew L, Lippincott Schwartz J, Zaal K. Fluorescent staining of subcellular organelles: ER, Golgi complex, and mitochondria. Curr Protoc Cell Biol. 2001;Chapter 4:Unit 4.4 pubmed publisher
    ..Fluorescent dyes and/or fluorescently labeled lipid derivatives can be used to identify ER, Golgi complex, and mitochondria. Specific conditions for labeling each of these compartments are described. ..
  4. Silva L, Coutinho A, Fedorov A, Prieto M. Conformation and self-assembly of a nystatin nitrobenzoxadiazole derivative in lipid membranes. Biochim Biophys Acta. 2003;1617:69-79 pubmed
    ..Therefore, we conclude that the amine group of nystatin is an essential requisite for the supramolecular organization/pore formation of this antibiotic. ..
  5. Ye F, Tao R, Cong W, Tian J, Liu Q. Utilization of fluorescence tracer in hyperinsulinemic-euglycemic clamp test in mice. J Biochem Biophys Methods. 2008;70:978-84 pubmed publisher
    ..All markers were reversed significantly by rosiglitazone treatment. The results indicate that with the fluorescence tracer 2-NBDG, the hyperinsulinemic-euglycemic clamp test can be used to estimate insulin sensitivity in vivo. ..
  6. Paulusma C, Folmer D, Ho Mok K, de Waart D, Hilarius P, Verhoeven A, et al. ATP8B1 requires an accessory protein for endoplasmic reticulum exit and plasma membrane lipid flippase activity. Hepatology. 2008;47:268-78 pubmed
    ..In the plasma membrane, ATP8B1 functions as a flippase for phosphatidylserine. Finally, CDC50A may be the potential beta-subunit or chaperone for ATP8B1 in hepatocytes. ..
  7. Higashi Y, Nakamura S, Fujii Y. Sensitive determination of 4-(4-bromophenyl)-4-hydroxypiperidine, a metabolite of bromperidol, in rat plasma by HPLC with fluorescence detection after pre-column derivatization using 4-fluoro-7-nitro-2,1,3-benzoxadiazole. Biol Pharm Bull. 2006;29:2479-82 pubmed
    ..Our method provides a sensitive procedure for determination of BPHP in rat plasma and is suitable for pharmacokinetic studies of BPHP after BRO administration. ..
  8. Darwish I, Al Majed A, Mahmoud A, Khalil N. New nonextractive and highly sensitive high-performance liquid chromatographic method for determination of paroxetine in plasma after offline precolumn derivatization with 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole. J AOAC Int. 2009;92:1349-55 pubmed
    ..The results demonstrated that the method would have a great value when it is applied in the therapeutic monitoring of PXT. ..
  9. Ball S, Bailey J, Stewart J, Vogels C, Westcott S. A fluorescent compound for glucose uptake measurements in isolated rat cardiomyocytes. Can J Physiol Pharmacol. 2002;80:205-9 pubmed
    ..040 nmol x min(-1) x 10(6) cells(-1), demonstrating the validity of this fluorescent compound for glucose uptake studies. ..

More Information


  1. Higashi Y, Nakamura S, Fujii Y. Sensitive determination of 4-(4-chlorophenyl)-4-hydroxypiperidine, a metabolite of haloperidol, in a rat biological sample by HPLC with fluorescence detection after pre-column derivatization using 4-fluoro-7-nitro-2,1,3-benzoxadiazole. Biomed Chromatogr. 2006;20:964-70 pubmed
    ..The method was simple and sensitive, useful for determination of CPHP in rat biological samples using as little as 200 microL of sample volume and could be applied for pharmacokinetic study. ..
  2. Erlichman J, Hewitt A, Damon T, Hart M, Kurascz J, Li A, et al. Inhibition of monocarboxylate transporter 2 in the retrotrapezoid nucleus in rats: a test of the astrocyte-neuron lactate-shuttle hypothesis. J Neurosci. 2008;28:4888-96 pubmed publisher
    ..Therefore, neurons seem to metabolize a combination of glucose and lactate (and other substances such as pyruvate) depending, in part, on the availability of each of these particular substrates. ..
  3. Zou C, Wang Y, Shen Z. 2-NBDG as a fluorescent indicator for direct glucose uptake measurement. J Biochem Biophys Methods. 2005;64:207-15 pubmed
    ..Our findings show that this technology has potential for applications in both medicine and research. ..
  4. Zhang H, Le Potier I, Smadja C, Zhang J, Taverna M. Fluorescent detection of peptides and amino acids for capillary electrophoresis via on-line derivatization with 4-fluoro-7-nitro-2,1,3-benzoxadiazole. Anal Bioanal Chem. 2006;386:1387-94 pubmed
    ..Peptides such as buccaline and beta-protein fragment 1-42 could also be derivatized using the developed in-capillary derivatization procedure. ..
  5. Zhang W, Lang C, Kaye D. Determination of plasma free 3-nitrotyrosine and tyrosine by reversed-phase liquid chromatography with 4-fluoro-7-nitrobenzofurazan derivatization. Biomed Chromatogr. 2007;21:273-8 pubmed
    ..067 +/- 0.050 nm/microm, respectively). In conclusion, we have developed a novel HPLC assay for NO(2)Tyr without MS detection that is applicable to clinical studies addressing the pathophysiology and importance of oxidative stress. ..
  6. Gupta V, Patwardhan G, Zhang Q, Cabot M, Jazwinski S, Liu Y. Direct quantitative determination of ceramide glycosylation in vivo: a new approach to evaluate cellular enzyme activity of glucosylceramide synthase. J Lipid Res. 2010;51:866-74 pubmed publisher
    ..These data indicate that this cell-based fluorescent method is direct, reproducible, and simple for assessing ceramide glycosylation. It is applicable to validate GCS activity in drug-resistant cancers and in other disorders. ..
  7. Bragg P, Hou C. Ligand-induced conformational changes in the Escherichia coli F1 adenosine triphosphatase probed by trypsin digestion. Biochim Biophys Acta. 1987;894:127-37 pubmed
    ..Since the digested enzyme lacked the epsilon subunit, a putative inhibitor of hydrolytic activity, a mechanism for the stimulation which involves loss or movement of this subunit is untenable. ..
  8. Ibuki Y, Suzuki A, Goto R. UVA irradiation induces energy-independent phospholipid-flip in mammalian plasma membrane. Photochem Photobiol. 2001;73:513-7 pubmed
    ..These findings indicated that UVA changes the flip-flop of phospholipids and that the cell membrane is a molecular and cellular target of UVA. ..
  9. Petrescu A, Gallegos A, Okamura Y, Strauss J, Schroeder F. Steroidogenic acute regulatory protein binds cholesterol and modulates mitochondrial membrane sterol domain dynamics. J Biol Chem. 2001;276:36970-82 pubmed
  10. Porras O, Loaiza A, Barros L. Glutamate mediates acute glucose transport inhibition in hippocampal neurons. J Neurosci. 2004;24:9669-73 pubmed
  11. Laquintana V, Denora N, Lopedota A, Suzuki H, Sawada M, Serra M, et al. N-benzyl-2-(6,8-dichloro-2-(4-chlorophenyl)imidazo[1,2-a]pyridin-3-yl)-N-(6-(7-nitrobenzo[c][1,2,5]oxadiazol-4-ylamino)hexyl)acetamide as a new fluorescent probe for peripheral benzodiazepine receptor and microglial cell visualization. Bioconjug Chem. 2007;18:1397-407 pubmed
    ..The obtained results indicate that compound 10 represents a new useful fluorescent probe for visualization of activated microglia and PBR. ..
  12. Elsing C, Gorski J, Boeker C, Stremmel W. Long-chain fatty acid uptake by skeletal myocytes: a confocal laser scanning microscopy study. Cell Mol Life Sci. 1998;54:744-50 pubmed
    ..Membrane hyper- and depolarization decreased uptake rates. This study demonstrates for the first time kinetic data from isolated myocytes with evidence for a carrier-mediated transport mechanism for long-chain fatty acids. ..
  13. Martin G, Atshaves B, Huang H, McIntosh A, Williams B, Pai P, et al. Hepatic phenotype of liver fatty acid binding protein gene-ablated mice. Am J Physiol Gastrointest Liver Physiol. 2009;297:G1053-65 pubmed publisher
    ..Taken together, these data support the hypothesis that L-FABP plays a role in physiological regulation of not only hepatic fatty acid metabolism, but also that of hepatic cholesterol. ..
  14. El Emam A, Hansen S, Moustafa M, El Ashry S, El Sherbiny D. Determination of lisinopril in dosage forms and spiked human plasma through derivatization with 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl) followed by spectrophotometry or HPLC with fluorimetric detection. J Pharm Biomed Anal. 2004;34:35-44 pubmed
    ..Interference from endogenous amino acids has been overcomed by using the solid phase extraction technique, the percentage recovery (n=6) was 101.6+/-3.35. ..
  15. Grubmeyer C, Gray W. A cysteine residue (cysteine-116) in the histidinol binding site of histidinol dehydrogenase. Biochemistry. 1986;25:4778-84 pubmed
    ..The presence of a cysteine residue in the histidinol binding site is consistent with models in which formation and subsequent oxidation of a thiohemiacetal occurs as an intermediate step in the overall reaction...
  16. Tyteca D, Van IJzendoorn S, Hoekstra D. Calmodulin modulates hepatic membrane polarity by protein kinase C-sensitive steps in the basolateral endocytic pathway. Exp Cell Res. 2005;310:293-302 pubmed
    ..This interference, although not affecting endocytosis as such, compromises cell polarity by impeding membrane trafficking from early endosomes to the apical membrane. ..
  17. Dimitriadis G, Maratou E, Boutati E, Psarra K, Papasteriades C, Raptis S. Evaluation of glucose transport and its regulation by insulin in human monocytes using flow cytometry. Cytometry A. 2005;64:27-33 pubmed
    ..Monocytes may be a valid model system to study the effects of insulin on glucose transport. Further, flow cytometry is suitable for this investigation and can be used as an alternative to radiotracer methods. ..
  18. Ito I, Watanabe S, Kirino Y. Mapping of odor-related neuronal activity using a fluorescent derivative of glucose. Neurosci Lett. 2006;398:224-9 pubmed
    ..Our results show that 2-NBDG is useful for mapping neuronal activity in vivo. ..
  19. Leira F, Louzao M, Vieites J, Botana L, Vieytes M. Fluorescent microplate cell assay to measure uptake and metabolism of glucose in normal human lung fibroblasts. Toxicol In Vitro. 2002;16:267-73 pubmed
    ..1 nM-1 microM) at different time intervals causes a decrease in intracellular glucose (40-50% over controls). Results obtained with okadaic acid are the starting point to evaluate application of the method to routine toxicity probes. ..
  20. Onal A, Sagirli O. Spectrophotometric and spectrofluorimetric methods for the determination of pregabalin in bulk and pharmaceutical preparation. Spectrochim Acta A Mol Biomol Spectrosc. 2009;72:68-71 pubmed publisher
    ..93% and 99.96% for spectrophotometric and spectrofluorimetric study, respectively. The suggested procedures could be used for the determination of PGB in pure and capsules being sensitive, simple and selective. ..
  21. Luo J, Fukuda E, Takase H, Fushinobu S, Shoun H, Wakagi T. Identification of the lysine residue responsible for coenzyme A binding in the heterodimeric 2-oxoacid:ferredoxin oxidoreductase from Sulfolobus tokodaii, a thermoacidophilic archaeon, using 4-fluoro-7-nitrobenzofurazan as an affinity label. Biochim Biophys Acta. 2009;1794:335-40 pubmed publisher
    ..K125A showed a large increase in the K(m) value for CoA and showed poor inactivation by NBD-F, compared with K173A and wild type StOFOR, indicating Lys125 in subunit-b is the critical residue that interacts with CoA. ..
  22. Williamson P, Christie A, Kohlin T, Schlegel R, Comfurius P, Harmsma M, et al. Phospholipid scramblase activation pathways in lymphocytes. Biochemistry. 2001;40:8065-72 pubmed
    ..These findings suggest that Ca(2+) and apoptosis operate through different pathways to activate the same scramblase. ..
  23. Roets E, Adams E, Muriithi I, Hoogmartens J. Determination of the relative amounts of the B and C components of neomycin by thin-layer chromatography using fluorescence detection. J Chromatogr A. 1995;696:131-8 pubmed
    ..The described method is much easier to perform than the official method. ..
  24. Fujioka K, Kozone I, Saito M, Matsuoka H. Rapid evaluation of the efficacy of microbial cell removal from fabrics. J Ind Microbiol Biotechnol. 2006;33:995-1002 pubmed
    ..After washing with a test detergent, no cell (<1) was detected. For this case, EMR was given by the formula: log(288/<1)=greater than 2.5. The imaging and cell count of a test fabric could be performed within 1 h. ..
  25. El Enany N. Spectrofluorimetric and spectrophotometric determination of gliclazide in pharmaceuticals by derivatization with 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole. J AOAC Int. 2003;86:209-14 pubmed
    ..The results were in good agreement with those obtained with the official and reference spectrophotometric methods. A proposal of the reaction pathway was presented. ..
  26. Massip C, Riollet P, Quemener E, Bayle C, Salvayre R, Couderc F, et al. Choice of different dyes to label tyrosine and nitrotyrosine. J Chromatogr A. 2002;979:209-15 pubmed
    ..A near infrared (780 nm) laser is used for NIR 780 derivatives. The UV-Visible limit of detection is 2.5 microM whereas it is in the range of 30 nM for LIF detection. ..
  27. Patkar K, Highsmith W, Aldrich J. Solid phase and solution synthesis of NvocLys(CO(CH2)5NH-NBD)OCH2CN, a trifunctional fluorescent lysine derivative. Amino Acids. 2009;36:203-7 pubmed publisher
    ..This synthetic strategy can be used for the preparation of a variety of amino acid derivatives with unusual alpha-amine and side chain functionalities. ..
  28. Loura L, Fedorov A, Prieto M. Exclusion of a cholesterol analog from the cholesterol-rich phase in model membranes. Biochim Biophys Acta. 2001;1511:236-43 pubmed
    ..It is concluded that, in particular, NBD-cholesterol is not a suitable cholesterol analog and its distribution behavior in phosphatidylcholine/cholesterol bilayers is in fact opposite to that of cholesterol. ..
  29. Tseng H, Gattolin S, Pritchard J, Newbury H, Barrett D. Analysis of mono-, di- and oligosaccharides by CE using a two-stage derivatization method and LIF detection. Electrophoresis. 2009;30:1399-405 pubmed publisher
    ..1 and 3%, respectively. The method was applied to measure sugars in nanoliter volume samples of phloem sap obtained by stylectomy from wheat and to honeydew samples obtained from aphids feeding from wheat and willow. ..
  30. Low M, Stutz P. Inhibition of the plasma glycosylphosphatidylinositol-specific phospholipase D by synthetic analogs of lipid A and phosphatidic acid. Arch Biochem Biophys. 1999;371:332-9 pubmed
    ..We propose that this diverse group of inhibitors all bind to a common site on GPI-PLD, the central hydrophobic cavity predicted by the beta-propeller model for integrin alpha subunits and GPI-PLD. ..
  31. Madrid E, Corchete P. Silymarin secretion and its elicitation by methyl jasmonate in cell cultures of Silybum marianum is mediated by phospholipase D-phosphatidic acid. J Exp Bot. 2010;61:747-54 pubmed publisher
    ..These results suggest that the enzyme PLD and its product PA mediate silymarin secretion to the medium of S. marianum cultures. ..
  32. Aoyama C, Tsunoda M, Funatsu T. Determination of selenomethionine by high-performance liquid chromatography-fluorescence detection coupled with on-line oxidation. Anal Sci. 2009;25:63-5 pubmed
    ..This simple and sensitive method should be useful for the determination of Se-Met in physiological samples, such as serum or urine. ..
  33. Hauck M, Kobler H. [Determination of cyclamate in complex matrix using HPLC after column derivatization with 4-fluoro-7-nitrobenzofurazan]. Z Lebensm Unters Forsch. 1990;191:322-4 pubmed
    ..The detection limit of cyclamate is 5 mg/kg foodstuff, with fluorescence detection 0.4 mg/kg. The recoveries are in the range of 88% to 104%. ..
  34. Weisiger R. Saturable stimulation of fatty acid transport through model cytoplasm by soluble binding protein. Am J Physiol. 1999;277:G109-19 pubmed
    ..However, slow interchange of fatty acid between the binding protein and membranes also appears to influence the transport rate in this model system. ..
  35. Pagano R, Sepanski M, Martin O. Molecular trapping of a fluorescent ceramide analogue at the Golgi apparatus of fixed cells: interaction with endogenous lipids provides a trans-Golgi marker for both light and electron microscopy. J Cell Biol. 1989;109:2067-79 pubmed
    ..Possible implications of these findings for metabolism and transport of (fluorescent) sphingolipids in vivo are also presented. ..
  36. Matsuoka H, Oishi K, Watanabe M, Kozone I, Saito M, Igimi S. Viable cell detection by the combined use of fluorescent glucose and fluorescent glycine. Biosci Biotechnol Biochem. 2003;67:2459-62 pubmed
    ..Thirty-five out of 41 strains showed marked uptake of 2NBDG but 6 strains were not able to take in 2NBDG. Five out of these 6 strains showed NBD-Gly uptake. ..
  37. Wu Y, Alexandrov K, Brunsveld L. Synthesis of a fluorescent analogue of geranylgeranyl pyrophosphate and its use in a high-throughput fluorometric assay for Rab geranylgeranyltransferase. Nat Protoc. 2007;2:2704-11 pubmed
    ..The procedure for the Rab geranylgeranyl transferase (RabGGTase) plate assay depends on the number of compounds in the screen but generally can be performed within a day. ..
  38. Kol M, van Laak A, Rijkers D, Killian J, de Kroon A, de Kruijff B. Phospholipid flop induced by transmembrane peptides in model membranes is modulated by lipid composition. Biochemistry. 2003;42:231-7 pubmed
    ..Finally, flop efficiency was strongly dependent on the phospholipid headgroup of the NBD-phospholipid analogue. Possible implications for transmembrane helix-induced flop in biomembranes in general are discussed. ..
  39. Lin W, Blanchette C, Ratto T, Longo M. Lipid asymmetry in DLPC/DSPC-supported lipid bilayers: a combined AFM and fluorescence microscopy study. Biophys J. 2006;90:228-37 pubmed
  40. Hrafnsdottir S, Nichols J, Menon A. Transbilayer movement of fluorescent phospholipids in Bacillus megaterium membrane vesicles. Biochemistry. 1997;36:4969-78 pubmed
    ..We conclude that Bacillus vesicles possess the ability to promote rapid transbilayer movement of phospholipids, and that the transport is probably protein (flippase)-mediated. ..
  41. Dekkers D, Comfurius P, van Gool R, Bevers E, Zwaal R. Multidrug resistance protein 1 regulates lipid asymmetry in erythrocyte membranes. Biochem J. 2000;350 Pt 2:531-5 pubmed
    ..This is the first report to show that MRP1 has a role in the maintenance of the outwards orientation of endogenous choline-containing phospholipids in the erythrocyte membrane...
  42. Maier O, Hoekstra D. Trans-Golgi network and subapical compartment of HepG2 cells display different properties in sorting and exiting of sphingolipids. J Biol Chem. 2003;278:164-73 pubmed
    ..The results indicate that in HepG2 cells sphingolipids are mainly sorted in the SAC membrane and that the release of SM from SAC and TGN is differentially regulated. ..
  43. Miller D, Graeff C, Droulle L, Fricker S, Fricker G. Xenobiotic efflux pumps in isolated fish brain capillaries. Am J Physiol Regul Integr Comp Physiol. 2002;282:R191-8 pubmed
  44. Boyanovsky B, Karakashian A, King K, Giltiay N, Nikolova Karakashian M. Uptake and metabolism of low density lipoproteins with elevated ceramide content by human microvascular endothelial cells: implications for the regulation of apoptosis. J Biol Chem. 2003;278:26992-9 pubmed
  45. Huang Q, Liang L, Wei T, Zhang D, Zeng Q. Purification and partial characterization of glutathione transferase from the teleost Monopterus albus. Comp Biochem Physiol C Toxicol Pharmacol. 2008;147:96-100 pubmed
    ..This high thermal stability of maGST could be related to the physiological adaptation of M. albus to high temperatures in tropical and subtropical environments. ..
  46. Kusano I, Kageyama A, Tamura T, Oda T, Muramatsu T. Enhancement of diphtheria toxin-induced apoptosis in Vero cells by combination treatment with brefeldin A and okadaic acid. Cell Struct Funct. 2001;26:279-88 pubmed
    ..Since a morphological change in the Golgi complex was observed in Vero cells treated with BFA and OA, modulation of the Golgi complex by these reagents may be partly responsible for enhanced apoptosis induction by diphtheria toxin. ..
  47. Louzao M, Espiña B, Vieytes M, Vega F, Rubiolo J, Baba O, et al. "Fluorescent glycogen" formation with sensibility for in vivo and in vitro detection. Glycoconj J. 2008;25:503-10 pubmed
    ..It could be used to record changes in glycogen stores in living cells and cell-free systems and opens the prospect of understanding the role of this important energy reserve under various physiological and pathophysiological conditions. ..
  48. Hu S, Li P. Micellar electrokinetic capillary chromatographic separation and fluorescent detection of amino acids derivatized with 4-fluoro-7-nitro-2,1,3-benzoxadiazole. J Chromatogr A. 2000;876:183-91 pubmed
    ..Furthermore, RSDs of migration time and peak height are better than 1% and 1.8%, respectively, again the smallest ever reported in the literature. ..
  49. Gadella B, Miller N, Colenbrander B, van Golde L, Harrison R. Flow cytometric detection of transbilayer movement of fluorescent phospholipid analogues across the boar sperm plasma membrane: elimination of labeling artifacts. Mol Reprod Dev. 1999;53:108-25 pubmed
    ..The described method enables, for the first time, assessment of changes in lipid asymmetry under fertilizing conditions...
  50. Zawadzki P, Slósarek G, Boryski J, Wojtaszek P. A fluorescence correlation spectroscopy study of ligand interaction with cytokinin-specific binding protein from mung bean. Biol Chem. 2010;391:43-53 pubmed publisher
  51. Bahrami G, Mohammadi B. Sensitive microanalysis of gabapentin by high-performance liquid chromatography in human serum using pre-column derivatization with 4-chloro-7-nitrobenzofurazan: Application to a bioequivalence study. J Chromatogr B Analyt Technol Biomed Life Sci. 2006;837:24-8 pubmed
    ..002-15 microg/ml. No interferences were found from commonly co-administrated antiepileptic drugs. The method was applied in a randomized cross-over bioequivalence study of two different gabapentin preparations in 24 healthy volunteers. ..
  52. Cai T, Guo Q, Wong B, Milot D, Zhang L, Wright S. Protein-disulfide isomerase is a component of an NBD-cholesterol monomerizing protein complex from hamster small intestine. Biochim Biophys Acta. 2002;1581:100-8 pubmed
    ..The p140 complex may thus contain an unidentified molecule(s) in addition to PDI that may contribute importantly to cholesterol monomerization. ..
  53. Roman Y, Alfonso A, Louzao M, Vieytes M, Botana L. Confocal microscopy study of the different patterns of 2-NBDG uptake in rabbit enterocytes in the apical and basal zone. Pflugers Arch. 2001;443:234-9 pubmed