Rapid cytotoxicity testing using a semi-automated protein determination on cultured cells

Shopsis, Charles; Eng, Ben

doi:10.1016/0378-4274(85)90176-6

Cited by 89 publications

(22 citation statements)

References 18 publications

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“…The method was carried out according to Shopsis and Eng [21] with modifications as follows. After 72 h exposure the medium was removed, and plates were transferred to a freezer (Ϫ80ЊC) and incubated for at least 6 h. Plates were removed from the freezer, and cells were allowed to thaw for 10 min at room temperature.…”

Section: Total Proteinsmentioning

confidence: 99%

An elisa assay for cytochrome P4501A in fish liver cells

Brüschweiler

Fent

Würgler

1996

Enviro Toxic and Chemistry

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Abstract-An enzyme-linked immunosorbent assay (ELISA) for measuring cytochrome P4501A (CYP1A) expression in vitro in fish hepatoma cells is described. Cells were cultured as monolayers in 96-microwell cell culture plates and exposed to polychlorinated biphenyl (PCB) congeners 77, 105, 153, and 169;, and ␤-naphthoflavone (BNF) for 3 d. Relative CYP1A protein content, CYP1A enzymatic activity, and total protein content were determined directly within the wells. At low concentrations of PCB 77, PCB 169, and 3-MC, the ethoxyresorufin-O-deethylase (EROD) activity was induced, but it was inhibited at high concentrations of these compounds. However, CYP1A protein content measured in an ELISA performed with intact cells increased monotonically in response to the concentration. No CYP1A induction was observed for PCB 105 and PCB 153. Because comparison between EROD activity and CYP1A amount gives information about the catalytic efficiency of CYP1A in the cells, this noncompetitive, solid-phase ELISA is recommended as a complementary method to the EROD assay. This novel ELISA method may be an accurate in vitro technique for a rapid and sensitive screening of CYP1A-inducible compounds.

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Section: Total Proteinsmentioning

confidence: 99%

An elisa assay for cytochrome P4501A in fish liver cells

Brüschweiler

Fent

Würgler

1996

Enviro Toxic and Chemistry

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“…Nas últimas décadas, vários grupos de pesquisadores vêm estudando e desenvolvendo métodos alternativos para determinação da irritação ocular (NOR-TH- ROOT et al, 1982;SHOPSIS;ENG, 1985;YANG;ACOSTA, 1994;VIAN et al, 1995;DE TORRES et al, 1997;TANI et al, 1999;ALVES et al, 2005;DEBBASCH et al, 2005;GERNER et al, 2005;LAGARTO et al, 2006;MEHLING et al, 2007;TAVASZI;BUDAI, 2007;ALVES et al 2008;MITJANS et al, 2008;ADRIAENS et al, 2008;ABREU, 2008;TAKAHASHI et al, 2008 Este estudo foi aprovado pela Comissão de Ética em Uso de Animais da Fiocruz (protocolos CEUA/Fiocruz P.0137-02 e P.0148-02).…”

Section: Introductionunclassified

Avaliação da irritabilidade ocular induzida por ingredientes de cosméticos através do teste de Draize e dos métodos HET-CAM e RBC.

Nóbrega¹,

Alves²,

Presgrave³

et al. 2009

Univ. Ci. Saúde

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“…Protein content of each well was determined in parallel to the pyruvate and H 2 O 2 assay, according to the methodology described by Shopsis and Eng [23]. The mesothelial cell monolayer was rinsed twice with Tris buffer saline; plates were incubated for 15 min at 37°C with 50 µl 0.1 N NaOH, and exposed to 200 µl of diluted dye reagent (1 part Coomassie Brilliant Blue G dye reagent (Sigma) – 3 parts distilled water) for 30 min.…”

Section: Methodsmentioning

confidence: 99%

Protective Effect of Pyruvate upon Cultured Mesothelial Cells Exposed to 2 m<i>M</i> Hydrogen Peroxide

Shostak

Gotloib²,

Kushnier³

et al. 2000

Nephron

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Rat peritoneal mesothelial cells in culture have the capability of generating hydrogen peroxide. Exposure of these cells to glucose-enriched, lactated-buffered fluids for peritoneal dialysis significantly increases the production of H₂O₂. Increased liberation of oxygen radicals also involves the risk of damaging the peritoneal membrane. Pyruvate being a natural oxidant scavenger abundantly present in mammalian cells, we hypothesized that its protective effects facing H₂O₂ can eventually be of relevance for the mesothelial monolayer of patients on long-term peritoneal dialysis. So far, we designed an experimental study in which rat peritoneal mesothelial cells in culture were exposed to 2 mM H₂O₂. Cell damage was estimated in terms of decreased capability of the mitochondrial dehydrogenases to reduce MTT. Addition of 2 mM sodium pyruvate to the medium prevented the negative effect of hydrogen peroxide. The MTT/protein values for the control group were 0.00357 ± 0.00075. The ratio after exposure to 2 mM H₂O₂ was 0.00217 ± 0.00028, whereas that detected in cells incubated in H₂O₂ plus pyruvate was 0.00325 ± 0.0082 (p < 0.05). These results indicate that pyruvate protected rat peritoneal mesothelial cells in culture against oxidant injury. These data are one more piece of evidence pointing at pyruvate as a potentially useful buffer for peritoneal dialysis solutions.

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Rapid cytotoxicity testing using a semi-automated protein determination on cultured cells

Cited by 89 publications

References 18 publications

An elisa assay for cytochrome P4501A in fish liver cells

An elisa assay for cytochrome P4501A in fish liver cells

Avaliação da irritabilidade ocular induzida por ingredientes de cosméticos através do teste de Draize e dos métodos HET-CAM e RBC.

Protective Effect of Pyruvate upon Cultured Mesothelial Cells Exposed to 2 m<i>M</i> Hydrogen Peroxide

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