Investigation of the bicinchoninic acid protein assay: Identification of the groups responsible for color formation

Wiechelman, Karen J.; Braun, Robert D.; Fitzpatrick, J. D.

doi:10.1016/0003-2697(88)90383-1

Cited by 649 publications

(351 citation statements)

References 7 publications

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“…The protein concentrations of the samples were determined using the Pierce BCA assay (Interchim, Asnières, France). 32 Results are expressed in RLU/ g protein ± standard deviation.…”

Section: Methodsmentioning

confidence: 99%

Minicircle: an improved DNA molecule for in vitro and in vivo gene transfer

et al. 1999

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Minicircles are a new form of supercoiled DNA molecule for enhancer/promoter. Comparing maximal differences, these nonviral gene transfer which have neither bacterial origin of minicircles gave 2.5 to 5.5 times more reporter gene replication nor antibiotic resistance marker. They are thus activity than the unrecombined plasmid in the NIH3T3 cell smaller and potentially safer than the standard plasmids line and rabbit smooth muscle cells. Moreover, injection in currently used in gene therapy. They were obtained in E.vivo into mouse cranial tibial muscle, or human head and coli by att site-specific recombination mediated by the neck carcinoma grafted in nude mice resulted in 13 to 50 phage integrase, which was used to excise the times more reporter gene expression with minicircles than expression cassette from the unwanted plasmid with the unrecombined plasmid or larger plasmids. Histosequences. We produced two minicircles containing the logical analysis in muscle showed there were more transluciferase or ␤-galactosidase gene under the control of fected myofibers with minicircles than with unrecombined the strong human cytomegalovirus immediate-early plasmid.

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“…The protein concentrations of the samples were determined using the Pierce BCA assay (Interchim, Asnières, France). 32 Results are expressed in RLU/ g protein ± standard deviation.…”

Section: Methodsmentioning

confidence: 99%

Minicircle: an improved DNA molecule for in vitro and in vivo gene transfer

et al. 1999

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“…Protein was measured by the bicinchoninic acid method [19] using a micro BCA protein assay reagent kit (Pierce) according to the manufacturer's instructions.…”

Section: Other Methodsmentioning

confidence: 99%

Identification and characterisation of a homologue of p64 in rat tissues

1996

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Previous work has suggested that the gene encoding p64, a component of a bovine kidney intraceHular chloride channel, may be a member of a gene family. We have raised a polyclonai antibody to an E. coli fusion protein which has sequence similarity to p64. Immunoblotting detected a protein in rat brain, kidney, liver and lung. In rat brain, the protein was enriched in cerebellar microsomal membranes. Western blot analyses of denaturing and blue native polyacrylamide gels indicated that the protein is a single non-disulphide-linked polypeptide chain with an apparent Mr of 43 kDa that contributes to a native protein complex with an apparent Mr of 130 kDa.

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“…Protein concentrations in the microsomal fractions were assessed using the bicinchoninic acid protein assay with serum albumin as a standard. 37 The concentration of CYP was estimated according to Omura and Sato 38 based on the absorption of the complex of reduced CYP with carbon monoxide. The content of CYP in human hepatic microsomes is shown in Table I.…”

Section: Preparation Of Microsomes and Assaysmentioning

confidence: 99%

Human hepatic and renal microsomes, cytochromes P450 1A1/2, NADPH:Cytochrome P450 reductase and prostaglandin H synthase mediate the formation of aristolochic acid‐DNA adducts found in patients with urothelial cancer

Stiborová

Frei

Hodek

et al. 2004

Intl Journal of Cancer

135

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Aristolochic acid (AA), a naturally occurring nephrotoxin and carcinogen, has been associated with the development of urothelial cancer in humans. Understanding which human enzymes are involved in AA activation and/or detoxication is important in the assessment of an individual's susceptibility to this plant carcinogen. Using the 32 P postlabeling assay, we examined the ability of microsomal samples from 8 human livers and from 1 human kidney to activate AAI, the major component of the plant extract AA, to metabolites forming adducts in DNA. Microsomes of both organs generated DNA adduct patterns reproducing those found in renal tissues from humans exposed to AA. 7-(deoxyadenosin-N 6 -yl)aristolactam I, 7-(deoxyguanosin-N 2 -yl)aristolactam I and 7-(deoxyadenosin-N 6 -yl)aristolactam II were identified as AA-DNA adducts formed from AAI by all human hepatic and renal microsomes. To define the role of human microsomal enzymes in the activation of AAI, we investigated the modulation of AAI-DNA adduct formation by cofactors and selective inhibitors of microsomal reductases, cytochrome P450 (CYP) enzymes, NADPH:CYP reductase and NADH:cytochrome b 5 reductase. We also determined whether the activities of CYP and NADPH:CYP reductase in different human hepatic microsomal samples correlated with the levels of AAI-DNA adducts formed by the same microsomal samples. On the basis of these studies, we attribute most of the activation of AAI in human hepatic microsomes to CYP1A2. In contrast to human hepatic microsomes, in human renal microsomes NADPH:CYP reductase is more effective in AAI activation. In addition, prostaglandin H synthase is another enzyme activating AAI in renal microsomes. The results demonstrate for the first time the potential of microsomal enzymes in human liver and kidney to activate AAI by nitroreduction.

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Investigation of the bicinchoninic acid protein assay: Identification of the groups responsible for color formation

Cited by 649 publications

References 7 publications

Minicircle: an improved DNA molecule for in vitro and in vivo gene transfer

Minicircle: an improved DNA molecule for in vitro and in vivo gene transfer

Identification and characterisation of a homologue of p64 in rat tissues

Human hepatic and renal microsomes, cytochromes P450 1A1/2, NADPH:Cytochrome P450 reductase and prostaglandin H synthase mediate the formation of aristolochic acid‐DNA adducts found in patients with urothelial cancer

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