Polarisation fluoroimmunoassay of gentamicin

Watson, R. A.; Landon, J.; Shaw, Elizabeth; Smith, David S.

doi:10.1016/0009-8981(76)90303-x

Cited by 70 publications

(21 citation statements)

References 7 publications

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“…No sample dilution is involved, as is the case with most immunoassays for gentamicin, such as the radioimmunoassay (4) and fluoroimmunoassay (8,9). Although this does mean that larger amounts of antiserum are needed, one rabbit can produce in 1 year enough antiserum for 100,000 tubes.…”

Section: Discussionmentioning

confidence: 99%

Rapid, Simple Enzyme Immunoassay for Gentamicin

Wills

Wise

1979

Antimicrob Agents Chemother

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Section: Discussionmentioning

confidence: 99%

Rapid, Simple Enzyme Immunoassay for Gentamicin

Wills

Wise

1979

Antimicrob Agents Chemother

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“…Column fractions were monitored for gentamicin content by enzyme-immunoassay (Emit, Syva-Biomerieux, France) and for fluorescein content by conventional fluorometry. The ratio of f1uorescein:gentamicin was 1: 1.4, and no unreacted FITC could be detected by TLC (Watson et al, 1976).…”

Section: Preparation and Uptake Of Fluorescein-labelled Gentamicinmentioning

confidence: 98%

Aminoglycoside antibiotics impair calcium entry but not viability and motility in isolated cochlear outer hair cells

Dulon

Zajic

Aran

et al. 1989

J of Neuroscience Research

124

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Cochlear outer hair cells have been well established as primary targets of the ototoxic actions of aminoglycoside antibiotics. These cells, isolated from the guinea pig cochlea and maintained in short-term culture, were used as a model for evaluating the acute effects of gentamicin on cell viability, depolarization-induced transmembrane calcium flux, and depolarization-induced motile responses. On the basis of morphology and fluorochromasia, the presence of extracellular gentamicin as high as 5 mM did not affect the viability of the cells for up to 6 hr, the longest time tested. Viable cells showed binding of fluorescently tagged gentamicin to their base but excluded the drug from their cytoplasm. In response to [K+]-depolarization, intracellular calcium levels (monitored with the fluorescent calcium-sensitive dye fluo-3) increased from a resting value of 218 +/- 102 nM to 2,018 +/- 1,077 nM concomitant with a cell shortening of 0.7% +/- 1.3%. The depolarization-induced calcium increase was apparently caused by calcium entry into the cell as it was inhibited by the calcium-channel blocker methoxyverapamil and prevented in the absence of extracellular calcium. Both gentamicin and neomycin blocked the [K+]-induced calcium increase at an IC50 of 50 microM. Despite the inhibition of calcium entry the ability of the outer hair cells to shorten under [K+]-depolarization was not impaired; in fact, cell shortening was even more pronounced in the absence of calcium influx (2.6% +/- 1.4%). This argues effectively against the existence of a calcium-dependent actomyosin-mediated component in [K+]-induced shape changes.(ABSTRACT TRUNCATED AT 250 WORDS)

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“…The reaction mixture was purified on a Sephadex column, with unreacted gentamicin being well separated from the fluorescein-labeled gentamicin. Polarization and quenching immunoassays (uide infra) were performed with the conjugate, and the results were comparable in sensitivity to bioassay and radioimmunoassay (25,26).…”

Section: Fluorescent Probesmentioning

confidence: 99%