Double-antibody solid-phase enzyme immunoassay for the detection of staphylococcal enterotoxin A

Saunders, George C.; Bartlett, M.L.

doi:10.1128/aem.34.5.518-522.1977

Cited by 82 publications

(28 citation statements)

References 17 publications

(7 reference statements)

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“…C6CM was tested for the presence of NGF, laminin, S-100, GFAP, neuron specific enolase (NSE), histone H I, and HMG 14 using ELISA, as well as immunoblots. ELISA was performed in microtiter plates according to Engvall and Perlman (1 972), with the modification that 2,2'-azinodi(3-ethylbenzthiazoline-6-sulfonate) was used as substrate (Saunders and Bartlett, 1977) instead ofp-nitrophenylphosphate. Plates were analyzed with an automatic ELISA reader at 492 nm (620 nm reference).…”

Section: Immunological Methodsmentioning

confidence: 99%

Neuronotrophic Factors Released by C6 Glioma Cells

Westermann

Hardung

Meyer

et al. 1988

Journal of Neurochemistry

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Glial cells have been shown previously to release factors that promote survival of central and peripheral neurons [neuronotrophic factors (NTFs)]. We have investigated the release of NTFs by C6 cells, a rat glioma cell line, under different modes of conditioning. Media conditioned in the presence or absence of serum [C6 cell conditioned media (C6CMs)] were analyzed using biological, biochemical, and immunological assays. We report that (a) nuclear and cytoskeletal proteins were not present in C6CMs, indicating that C6CM proteins result from release by C6 cells rather than from cell death; (b) C6CM contained 1-3 micrograms protein/ml, corresponding to a secretion rate of about 0.5 pg protein per cell and day; (c) C6CM contained the neurite-promoting factor laminin and low amounts of nerve growth factor; (d) the presence of fetal calf serum in the culture medium was essential for synthesis and release of NTFs; and (e) our C6CM contained at least three NTFs differing by their temporal secretory patterns and three NTFs differing by biochemical properties, indicating that C6 cells produce and secrete six different NTFs. Within these, nerve growth factor seems to be the only established NTF.

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Section: Immunological Methodsmentioning

confidence: 99%

Neuronotrophic Factors Released by C6 Glioma Cells

Westermann

Hardung

Meyer

et al. 1988

Journal of Neurochemistry

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“…One hundred p1 of substrate containing 50 mM citrate buffer (pH 4.0), 0.4 mM ABTS, 0.06 mM H202 were then added to each vial. Following development of color (approximately 5 mid, the reaction was terminated by adding 1OOpl HF-edetic acid stopping reagent (Saunders 1979;Saunders and Bartlett 1977). Absorbance was determined in a Beckman DU spectrophotometer equipped with a flow cell (Altex Model 1541, a peristaltic pump (Pharmacia P-31, and a strip chart recorder (Heath Schlumberger Model EU-205-11) .…”

Section: Determination Of Antiserum Titer By Elisa Methodsmentioning

confidence: 99%

Production and Characterization of Antibody Against Aflatoxin G1

Chu

Steinert

Gaur

1985

Journal of Food Safety

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Antibody against aflatoxin G1 (AFG1) was obtained from rabbits after immunizing the animals with AFG1 hemiacetal (AFG2a) conjugated to bovine serum albumin. A direct heterogenous ELISA in which AFG2a was conjugated to horseradish peroxidase was used for monitoring the antibody titers and for toxin detection. Competitive ELISA assay revealed that the antibody was most specific for AFG2a and least for AFB2. The relative cross‐reactivity of this antiserum with aflatoxins G2a, G2, G1, M1, E1 and B2 was found to be 1, 7, 13, 47, 48, and 63, respectively. The lower detection limits for detection of AFG1 after derivatizing to AFG2a was around 15–25 pg per assay.

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“…Enzyme Linked Immunosorbent Assay (ELISA) (Notermans et al 1978;Simon and Terplan 1977;Fey 1978), Radioimmunoassay (RIA) (Miller e t al. 1978;Lindroth and Niskanen 1977;Pober and Silverman 1977;Robern et al 1978), Enzyme Immuno Assay (EIA) (Saunders and Bartlett 1977), Homogeneous Enzyme Immune Assay (Morita and Woodburn 1978) and Reversed Passive Hemagglutination Assay (RPHA) (Shinagawa et al 1977b). Some of these methods can be used after a greatly simplified extraction procedure.…”

Section: Introductionmentioning

confidence: 99%

“…Some of these methods can be used after a greatly simplified extraction procedure. To our know-ledge, however, most of these methods have so far only been applied to the quantification of purified toxin, that had been added to food (Robern et al 1978), or even t o food extracts (Lindroth and Niskanen 1977;Pober and Silverman 1977;Saunders and Bartlett 1977).…”

Section: Introductionmentioning

confidence: 99%

PREVENTION OF CROSS‐REACTIONS IN THE ENZYME LINKED IMMUNOSORBENT ASSAY (ELISA) FOR THE DETECTION OF STAPHYLOCOCCUS AUREUS ENTEROTOXIN TYPE B IN CULTURE FILTRATES AND FOODS

Koper

Hagenaars

Notermans

1980

Journal of Food Safety

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Investigations were performed to avoid cross‐reactions in the enzyme linked immunosorbent assay, using the sandwich technique, for detection of Staphylococcus aureus enterotoxin type B. Non‐specific reactions can be caused by cross‐reacting antigens and by Protein A, produced by S. aureus. The former reactions can be prevented by adsorption with culture filtrate of non‐toxin type B producing strains. The latter reaction is caused by the binding of Protein A to the Fc fragments of the IgG antibodies. Interference by Protein A was completely eliminated by using the F(ab')2 fragments of the IgG antibodies. ELISA experiments in which these purified F(ab')2 fragments were used resulted in a highly specific detection of entertoxin type B, both in culture filtrates and in foods.

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Double-antibody solid-phase enzyme immunoassay for the detection of staphylococcal enterotoxin A

Cited by 82 publications

References 17 publications

Neuronotrophic Factors Released by C6 Glioma Cells

Neuronotrophic Factors Released by C6 Glioma Cells

Production and Characterization of Antibody Against Aflatoxin G1

PREVENTION OF CROSS‐REACTIONS IN THE ENZYME LINKED IMMUNOSORBENT ASSAY (ELISA) FOR THE DETECTION OF STAPHYLOCOCCUS AUREUS ENTEROTOXIN TYPE B IN CULTURE FILTRATES AND FOODS

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