Quantitation of human interleukin-1β with polyclonal antibodies

Gaffney, Edwin V.; Koch, G; Tsai, Shiow-Chuan J.; Lingenfelter, Susan E.; Piscitelli, Joseph

doi:10.1016/0022-1759(87)90160-8

Cited by 10 publications

(3 citation statements)

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“…Antibodies produced against a recombinant product may not bind to the naturally occurring protein. Therefore, antibodies must be chosen that recognize epitopes shared by the recombinant and natural cytokines (Gaffney et al, 1987;Abrams and Pearce, 1988;Sato et al, 1986).…”

Section: Natural and Recombinant Cytokinesmentioning

confidence: 99%

“…All antibodies must be rigorously screened by western blotting for cross-reactivity and non-specific binding to other molecules. Both polyclonal and monoclonal antibodies have been successfully used in cytokine immunoassays (Wadhwa et al, 1990;DeForge and Remick, 1991;Gaffney et al, 1987).…”

Section: Natural and Recombinant Cytokinesmentioning

confidence: 99%

“…The capture ELISA requires two antibodies that bind two separate epitopes so that the capture antibody and the enzyme-linked antibody can bind the cytokine simultaneously. These requirements can be satisfied with one or two polyclonal (Gaffney et al, 1987), two monoclonal (Schumacher et al, 1988;Andersson et al, 1989) or a combination of polyclonal and monoclonal antibodies (Wadhwa et al, 1990;Ko et al, 1992). Most commercially available cytokine ELISA "ki ts" use the capture or sandwich format and they have a detection limit of approximately 10 picograms.…”

Section: Enzyme-linked Immunosorbent Assaymentioning

confidence: 99%

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Cloning bovine cytokine cDNA fragments and measuring bovine cytokine mRNA using the reverse transcription-polymerase chain reaction

Hutchinson

Stevens

Olsen

1994

Veterinary Immunology and Immunopathology

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Bovine cytokine-specific primers and the reverse transcription-polymerase chain reaction (RT-PCR) were used to clone cDNA fragments that were specific for bovine IL-1 alpha, IL-1 beta, IL-2, and IFN-gamma. Specificity of the cDNA fragments was verified by sequence analysis based on known bovine IL-1 alpha, IL-1 beta, IL-2, and IFN-gamma gene sequences. In addition, RT-PCR was used to monitor cytokine mRNA expression in concanavalin A (Con A) and lipopolysaccharide (LPS)-stimulated bovine peripheral blood mononuclear cells (PBMC), and the results were compared with those obtained by measuring PBMC cytokine secretion using biologic assays. IL-1 activity in LPS-stimulated PBMC cultures was similar at 12 h and 24 h, although the activity decreased by approximately 40% at 48 h. IL-2 and IFN-gamma activity in supernatants of Con A-stimulated PBMC cultures was low at 12 h and reached maximum levels at 48 h. RT-PCR transcript analysis detected an increase in IL-1 alpha, IL-1 beta, IL-2, and IFN-gamma mRNA expression that was usually correlated with the detection of these soluble cytokines by the bioassays. These results indicate that RT-PCR is a sensitive and effective method of obtaining cDNA probes and that this technique can be used to monitor bovine cytokine mRNA expression.

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Section: Natural and Recombinant Cytokinesmentioning

confidence: 99%

Section: Natural and Recombinant Cytokinesmentioning

confidence: 99%