Assay to measure CD59 mutations in CHO AL cells using flow cytometry
| العنوان: | Assay to measure CD59 mutations in CHO AL cells using flow cytometry |
|---|---|
| المؤلفون: | Chang-Uk Lim, Michael H. Fox, Carley D. Ross |
| المصدر: | Cytometry Part A. :85-90 |
| بيانات النشر: | Wiley, 2005. |
| سنة النشر: | 2005 |
| مصطلحات موضوعية: | Methylnitronitrosoguanidine, Time Factors, Histology, medicine.drug_class, DNA Mutational Analysis, Mutant, Mutagenesis (molecular biology technique), CD59 Antigens, CHO Cells, Cell Separation, Hybrid Cells, Biology, Monoclonal antibody, medicine.disease_cause, Sensitivity and Specificity, Article, Pathology and Forensic Medicine, Flow cytometry, chemistry.chemical_compound, Clastogen, Cricetinae, medicine, Animals, Humans, Mutation, medicine.diagnostic_test, Mutagenicity Tests, Chromosomes, Human, Pair 11, Chinese hamster ovary cell, Antibodies, Monoclonal, Cell Biology, Flow Cytometry, Molecular biology, chemistry, Gamma Rays, Mutagenesis, Calibration |
| الوصف: | Background A sensitive mammalian cell mutation assay was developed previously using a Chinese hamster ovary cell line (CHO AL) that stably incorporates human chromosome 11. The assay measures mutations in the CD59 gene on chromosome 11 but it requires the use of rabbit complement and colony growth for mutant selection. We have developed a more rapid flow cytometry-based mutation assay with CHO AL cells that uses monoclonal antibodies against CD59 to detect mutants and does not require colony formation. Methods CHO AL cells were treated with γ-radiation or N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) and then allowed to grow for various times for mutant expression. Cells were labeled with monoclonal antibodies against CD59 and analyzed by flow cytometry. Results Negative and positive populations were separated by over 100-fold. Mixing various proportions of CD59-positive and -negative cells demonstrated that the assay is highly linear (r2 = 0.9999) and sensitive ( |
| تدمد: | 1552-4930 1552-4922 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::fa04e715b53091f22722cfb1b4aabb12Test https://doi.org/10.1002/cyto.a.20168Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....fa04e715b53091f22722cfb1b4aabb12 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15524930 15524922 |
|---|