Abstract：
:A method for quantitative RT-PCR using ELISA detection was developed and applied to the quantitation of IL-1 beta mRNA in clinical samples. To compensate for the 'tube effect' of RT-PCR, a synthetic RNA, pRSET RNA, was added to sample solutions as an internal standard and co-amplified with IL-1 beta mRNA. Sense primers for IL-1 beta and pRSET were labeled with digoxigenin and FITC, respectively, while anti-sense primers for both were labeled with biotin. Double-stranded PCR products were captured by two solid-phase pipettetips. One was coated with an anti-digoxigenin antibody and another with an anti-FITC antibody, and sandwiched by avidin-urease, and their activities were measured by coupling them with a pH-FET in a pH-measuring cell containing urea solution. The ratio of the signal intensity for IL-1 beta to that for pRSET was used to quantify the concentration of IL-1 beta mRNA. A calibration curve was obtained by using a known amount of AW 109 RNA as an external standard of IL-1 beta RNA. It was found that 10(2)-10(6) copies of IL-1 beta mRNA were measurable by the present method. Expression levels of IL-1 beta mRNA in clinical samples, such as monocytes of peripheral blood or synovial cells from patients with RA or OA, were determined.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Tsuruta H,Matsui S,Oka K,Namba T,Shinngu M,Nakamura Mdoi
10.1016/0022-1759(94)00321-msubject
Has Abstractpub_date
1995-03-27 00:00:00pages
259-64issue
2eissn
0022-1759issn
1872-7905pii
002217599400321Mjournal_volume
180pub_type
杂志文章abstract：:To maximize safety in the setting of an increasing number of requests for flow cytometric analysis of specimens potentially contaminated with the AIDS retrovirus, we evaluated some commonly used fixatives for their ability to inactivate the infectious potential of the virus. We found that both formaldehyde (0.37% v/v)...
journal_title：Journal of immunological methods
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journal_title：Journal of immunological methods
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journal_title：Journal of immunological methods
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90317-z
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journal_title：Journal of immunological methods
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journal_title：Journal of immunological methods
pub_type： 杂志文章
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journal_title：Journal of immunological methods
pub_type： 杂志文章
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journal_title：Journal of immunological methods
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journal_title：Journal of immunological methods
pub_type： 杂志文章
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journal_title：Journal of immunological methods
pub_type： 杂志文章
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(80)90058-7
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journal_title：Journal of immunological methods
pub_type： 杂志文章
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journal_title：Journal of immunological methods
pub_type： 杂志文章
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90162-9
更新日期：1991-04-25 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(01)00447-1
更新日期：2001-11-01 00:00:00
abstract：:A sensitive time-resolved fluoroimmunoassay of nerve growth factor (NGF) has been developed. The method is based on the unique property of the lanthanides for delayed fluorescence, which reduces substantially the endogenous fluorescence of biological substances, because the excitation of the sample and detection of th...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90334-7
更新日期：1989-08-15 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(80)90336-1
更新日期：1980-01-01 00:00:00
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journal_title：Journal of immunological methods
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journal_title：Journal of immunological methods
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journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
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更新日期：1996-09-13 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
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journal_title：Journal of immunological methods
pub_type： 杂志文章
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更新日期：1995-06-14 00:00:00
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journal_title：Journal of immunological methods
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更新日期：2017-02-01 00:00:00
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journal_title：Journal of immunological methods
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更新日期：2019-08-01 00:00:00
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journal_title：Journal of immunological methods
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更新日期：2003-04-01 00:00:00
abstract：:Giant liposomes (mean diameter 5.5 microns) composed of egg phosphatidylcholine or distearoyl phosphatidylcholine, phosphatidyl glycerol, cholesterol and triolein were prepared by a double emulsion technique. They were then mixed with model particulate (killed Bacillus subtilis, and killed Bacille Calmette-Guérin) and...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90368-h
更新日期：1993-12-03 00:00:00
abstract：:This paper shows how to obtain accuracy and efficiency in an ELISA analysis by allocating the wells on a 96-well microplate between calibration and determination of unknowns, and by choosing the known concentrations for calibration. The method also can determine how much is lost in precision by using a convenient but ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90036-u
更新日期：1990-09-14 00:00:00
abstract：:CD4(+) T cells play a central role in regulating the immune response. Their effector function is commonly assessed by their capacity to secrete cytokines detected by ELISPOT and intracellular cytokine staining. However, one aspect of their effector function that is often overlooked is their ability to help activation ...
journal_title：Journal of immunological methods
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更新日期：2013-01-31 00:00:00
