Establishment of a highly sensitive enzyme-linked immunosorbent assay for interleukin-1α employing a fluorogenic substrate

Mukaida, Naofumi; Kimura, Tadashi; Ko, Y C; Kawai, Tadashi

doi:10.1016/0022-1759(88)90006-3

Cited by 8 publications

(2 citation statements)

References 19 publications

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“…Myosin extracts were diluted with 10 mM Tris-HC1 buffer, pH 7.4 containing 0.5 M NaC1, 0.3% gelatin and 0.05% Tween-20, and then 100 gl aliquots were incubated on the mAb-coated microstrips for 1 h at room temperature. The myosins bound to the solid-phase mAb were detected by using biotin-conjugated BBM4 as a detector antibody, and followed by ~-galactosidase-conjugated streptavidin (Boehringer Mannheim, Germany) and 4-methylumbelliferyl-~-D-galactopyranoside (New Brunswick, UK) as reagents for production of fluorescent products [6]. The fluorescence intensity (EX, 360 nm; EM, 450 rim) was measured with a microplate reader (MTP-100F, CORONA, Japan).…”

Section: Enzyme-linked Immunosorbent Assay (Elisa)mentioning

confidence: 99%

Brain tissue identification based on myosin heavy chain isoforms

et al. 1995

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Non-muscle tissues contain 3 myosin heavy chain (MHC) isoforms; MIIA, MIIB1 and MIIB2. MIIA is a non-muscle type isoform distributed in all non-muscle tissues and smooth-muscle, while MIIB1 and MIIB2 are brain-type isoforms distributed mainly in neuronal tissues. The ratio of MIIA and MIIB (A/B ratio) differs between tissues, suggesting that this ratio may be a useful marker for tissue identification. To apply the A/B ratio for tissue identification in forensic practice, we developed a highly sensitive ELISA for quantification of each MHC isoform. At least 100 pg of both MHC isoforms could be detected by the present method. Analysis of the A/B ratio of the cerebrum, cerebellum, liver, kidney, spleen and andrenal gland by the present method indicated that the A/B ratio of the brain tissue (< 0.5) was quite different from other tissues (> 3.0). The A/B ratio could be determined from at least 8 micrograms of fresh tissue sample and 0.1 mg of dried tissue sample stored for 1 month at room temperature. Therefore, the A/B ratio seems to be an excellent marker for identification of the brain tissue.

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Section: Enzyme-linked Immunosorbent Assay (Elisa)mentioning

confidence: 99%

Brain tissue identification based on myosin heavy chain isoforms

et al. 1995

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“…Several monoclonal and polyclonal antibodies have been produced to purified native IL-1, rIL-1, or peptide fragments of IL-1 (Mukaida et al 1988, Stya et al 1988) and these have been utilized in the design of commercially available immunoassays. The first radioimmunoassay for IL-1 measured the levels of the endogenous pyrogen produced by purified human monocytes following exposure to staphylococci (Dinarello et al 1977).…”

Section: Il-1 Assaysmentioning

confidence: 99%

Antibodies and their Role in Interleukin‐1 Research

Gaffney

Anderson

Pfanenstiel

et al. 1991

Immunological Reviews

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The preceding paragraphs describe several uses of specific antibodies to IL-1. Standard radioimmunoassays and ELISA procedures were developed with demonstrated utility in studies attempting to quantitate the cytokine in cell culture fluids. In part, antibody-based assays circumvent the problems associated with bioassays. Bioassays are difficult to perform, often lack specificity due to the presence of competing cytokines and are insensitive when used with complex biological fluids due to non-specific protein binding and the presence of IL-1 antagonists. Thus, bioassays have not been broadly applicable with specimens in a clinical setting. In contrast, monoclonal antibodies and polyclonal antisera have provided a means of identifying functionally important regions of the IL-1 molecule, monitoring the post-translational processing of IL-1 which may include biologically inactive moieties, and developing new assays as illustrated by cell blot procedures.

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