Abstract:
:Using in vitro established tumour cell lines attempts were made to assess the suitability of tetrazolium salt reduction (MTT) assay to replace the conventional radioactive base techniques for measuring cell proliferation and cell killing. The optimum conditions for MTT loading time, concentration of MTT and the time for colour development were found to be 4 h, 5 mg/ml and 30 min respectively, conditions which were used for subsequent experiments. Analysis of the correlation between increasing cell numbers and optical densities (OD) showed a direct relationship with correlation of coefficient values of r > 0.98 and 10,000 cells/well was found to provide an accurate ODs for a wide variety of cell types. The accuracy of replicate readings of the assay was investigated by setting a wide range of cell numbers and the variation among seven replicates was calculated and found to be less that 6% of the mean values. The reproducibility of the assay for two cell lines was tested using the lines on four different occasions. The ODs for Jar and Fen cell lines were 0.80 +/- 0.01, 0.82 +/- 0.02, 0.90 +/- 0.02, 0.79 +/- 0.05 and 0.56 +/- 0.01, 0.58 +/- 0.03, 0.60 +/- 0.02 and 0.61 +/- 0.02 respectively giving maximum variation of less than 11% of mean on repeated testings. Comparison between the results of MTT with 3H-Tdr or 51Cr release assays showed a high degree of correlation over a wide range of cell numbers and cell types. The r values between the results of MTT with 3H-Tdr (for cell number ranging from 1.8 to 60 x 10(3)/well) or 51Cr release assays (for E/T ratios of between 5:1 and 40:1) were 0.89 (p = 0.001) and 0.96 (p < 0.03) respectively. These results demonstrate that it is possible to use the MTT assay interchangeably with radioactive base techniques to measure cell proliferation and cytotoxicity. The ease of its execution, safety and its suitability for analysing as few as 3000 cells makes this method a serious contender for replacing the conventional radioactive techniques.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Hussain RF,Nouri AM,Oliver RTdoi
10.1016/0022-1759(93)90012-vsubject
Has Abstractpub_date
1993-03-15 00:00:00pages
89-96issue
1eissn
0022-1759issn
1872-7905pii
0022-1759(93)90012-Vjournal_volume
160pub_type
杂志文章abstract::The majority of vaccines and several treatments are administered by intramuscular injection. The aim is to engage and activate immune cells, although they are rare in normal skeletal muscle. The phenotype and function of resident as well as infiltrating immune cells in the muscle after injection are largely unknown. W...
journal_title:Journal of immunological methods
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abstract::An enzyme-linked immunosorbent assay (ELISA) for determination of antibodies against the zona pellucida was developed and compared with the already available indirect immunofluorescence (IIF) technique. Sera from 100 women with explained and unexplained infertility were screened for the presence of autoantibodies to t...
journal_title:Journal of immunological methods
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abstract::A fluorescence quenching method, using trypan blue, is described for quantifying the ingestion of either glutaraldehyde-fixed sheep red blood cells or antibody coated glutaraldehyde-fixed sheep red blood cells by murine peritoneal macrophages. This method is based on the observations that glutaraldehyde-fixed red bloo...
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(86)90438-2
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abstract::The detection of specifically internal immunoglobulin by staining of fixed preparations of lymphoid cells with appropriate fluorescein-conjugated antisera can be hampered by binding of the conjugates to membrane-bound immunoglobulin. Without preventing staining through membrane-bound immunoglobulin, it is impossible ...
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(85)90223-6
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journal_title:Journal of immunological methods
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abstract::Among the many methods currently available for quantifying mRNA transcript abundance, reverse transcription-polymerase chain reaction (RT-PCR) has proved to be the most sensitive. Recently, several protocols for real-time relative RT-PCR using the reporter dye SYBR Green I have appeared in the literature. In these met...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(03)00103-0
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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
doi:10.1016/s0022-1759(98)00086-6
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journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2006.05.008
更新日期:2006-07-31 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)90252-5
更新日期:1980-01-01 00:00:00
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journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(86)90231-0
更新日期:1986-11-20 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
doi:10.1016/0022-1759(91)90154-8
更新日期:1991-08-09 00:00:00
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journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2010.06.007
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journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(87)90191-8
更新日期:1987-06-26 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
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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(95)00103-h
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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:1997-03-10 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
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journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(97)90008-9
更新日期:1977-01-01 00:00:00
abstract::Protein phosphorylation is a major molecular mechanism by which cellular function is regulated. In order to accomplish rapid and specific biochemical changes via phosphorylation, the activity of a protein kinase must be dynamically regulated. Historically, the activity of each protein kinase has been analyzed using a ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
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更新日期:2000-12-01 00:00:00