Производитель может поднять цены — запросите коммерческое предложение сейчас, и мы зафиксируем за вами текущую цену.
-
Привезем под заказ -
Срок поставки с завода 6-8 недель
5-Bromo-2-deoxy-uridine Labeling and Detection Kit III 11444611001
Гарантия 1 год
- Код:
- 11444611001
- Производитель:
- Roche
- Производитель:
- Roche
General description
Proliferation in cell populations may be studied by incorporating the radioisotope [3H]-thymidine into cellular DNA. The amount of radioactive thymidine incorporated is determined by scintillation counting. Alternatively , 5-bromo-2-deoxy-uridine may be incorporated into cellular DNA. The amount of BrdU incorporated is determined by a standard ELISA protocol, which involves "tagging" the incorporated nucleotide with an anti-BrdU antibody. 96-well microplate cell ELISA for the detection of 5-bromo-2-deoxy-uridine (BrdU) incorporated into cellular DNA. Nonradioactive alternative for [3H]-thymidine based DNA synthesis and cell proliferation assays.Contents
- BrdU Labeling Reagent, 1,000x concentrated
- Washing Buffer concentrate, 10x concentrated
- Incubation Buffer
- Nucleases
- Anti-BrdU-POD, Fab fragments
- Substrate Buffer
- ABTS Substrate
- Substrate Enhancer
Specificity
Anti-BrdU-peroxidase, Fab fragments, specifically bind to 5-bromo-2′-deoxy-uridine incorporated into DNA. It shows no cross-reactivity with any endogenous cellular components, such as thymidine or uridine.
Application
The kit is used for the quantitative determination of BrdU incorporated into cellular DNA using a 96-well microplate cell ELISA format.5-Bromo-2-deoxy-uridine Labeling and Detection Kit III has been used in cell proliferation assay, BrdU assay and proliferation assay.
Features and Benefits
- Safer, since the kit does not use radioisotopes.
- Accurate, since results generated with this assay strongly correlate to those obtained with the [3 H]-thymidine method. (See Figure 2 below.)
- Sensitive. This assay and the [3 H]-thymidine assay are equally sensitive. (See Figure 2 below.)
- Fast, since results can be read with a multi-well ELISA reader, allowing simultaneous processsing of a large number of samples
- Easy, since the assay uses a standard cell ELISA protocol.
- Economical, since the assay requires no expensive equipment or additional reagents (e.g ., scintillation fluid).
Packaging
1 kit containing 8 components.
Principle
Cells cultured in a 96-well microplate are incubated with BrdU (see Performance). The labeled cells are fixed with ethanol. Prior to incubation with a monoclonal antibody to BrdU, DNA is partially digested with nucleases to allow the antibody to access BrdU. Next, the anti-BrdU antibody [labeled with peroxidase (POD)] is added. Finally, the POD substrate ABTS is added. POD catalyzes the cleavage of ABTS, producing a colored reaction product. The absorbance of the samples (at approximately 405 nm) is determined with a standard microplate (ELISA) reader.
Preparation Note
Working solution: BrdU labeling solution
Dilute BrdU labeling reagent 1 : 90 with sterile PBS or culture medium (resulting concentration: 111 µM BrdU) [e.g., for one 96-well microplate containing 100 µl medium per well, dilute 12 µl BrdU labeling reagent with 1.068 ml sterile PBS].
Note: The BrdU labeling solution should be prepared freshly before use.
Washing buffer
Dilute washing buffer concentrate (10x) 1 : 10 with double-dist. water [e.g., for one 96-well microplate dilute 9 ml washing buffer concentrate (10x) with 81 ml double-dist. water].
Note: If precipitates in Washing buffer, 10x conc. are visible, please incubate the bottle for 10 minutes at 37 °C in a water bath before you prepare Solution II.
Washing buffer is used to:
- Prepare the anti-BrdU-peroxidase, working solution
- Wash cells after incubation with anti-BrdU-peroxidase
Incubation buffer
- Ready-to-use
- Used to dilute the nucleases
Nucleases, stock solution
Reconstitute the nucleases in 1.3 ml double-dist. water containing 50% glycerol (w/v).
Nucleases, working solution
Dilute nucleases, stock solution, 1 : 100 with incubation buffer (e.g., for one 96-well microplate dilute 100 µl nucleases, stock solution, with 9.9 ml incubation buffer).
Anti-BrdU-peroxidase, Fab fragments, stock solution
Dissolve anti-BrdU-peroxidase, Fab fragments in 1.25 ml double-dist. water (final concentration: 20 U/ml).
Anti-BrdU-peroxidase, Fab fragments, working solution
Prepare anti-BrdU-peroxidase, Fab fragments, working solution shortly before use. Dilute anti-BrdU-peroxidase, Fab fragments, stock solution 1 : 100 with washing buffer supplemented with 10 mg/ml BSA (bovine serum albumin), [e.g., for one 96-well microplate dilute 100 µl anti-BrdU-peroxidase, Fab fragments, stock solution, with 9.9 ml PBS and BSA (final concentration: 200 mU/ml)].
Peroxidase substrate
Dissolve the ABTS powder in substrate buffer and stir at 15 to 25 °C to obtain a clear solution.
Peroxidase substrate containing substrate enhancer
If a low signal is expected, take an appropriate aliquot of substrate solution and add substrate enhancer , 1 mg/ml and dissolve by stirring for 15 minutes at 15 to 25 °C (e.g., for one 96-well microplate dissolve 10 mg substrate enhancer in 10 ml peroxidase substrate)
Note: The substrate solution containing substrate enhancer is stable for only 4 hours and should, therefore, be freshly prepared before use.
Storage conditions (working solution): BrdU labeling reagent
The undiluted BrdU labeling reagent (1000x) is stable at 2 to 8 °C for 6 months. It is stable stored in aliquots at -15 to -25 °C.
Washing buffer
Stable at 2 to 8 °C for 3 months.
Incubation buffer
Stable at 2 to 8 °C until the expiration date printed on the label
Nucleases, stock solution
Stable at -15 to -25 °C for 6 months.
Nucleases, working solution
must be prepared shortly before use.
Anti-BrdU-Peroxidase, Fab fragments, stock solution
Stable at 2 to 8 °C for 6 months. For long term storage it is recommended to store the solution in aliquots at -15 to -25 °C.
Anti-BrdU-Peroxidase, Fab fragments, working solution
must be prepared freshly before use.
Peroxidase substrate
Stable at 2 to 8 °C for 2 months when stored protected from light.
Peroxidase substrate containing substrate enhancer
must be prepared freshly before use
Other Notes
For life science research only. Not for use in diagnostic procedures.Параметры
| usage | sufficient for ≤1,000 tests |
| Quality Level | 100 |
| specific activity | 10000 (Nuclease activity (vial 4)) |
| Торговая марка | Roche |
| shipped in | wet ice |
| storage temp. | 2-8°C |
Safety Information
| pictograms | GHS07 |
| signalword | Warning |
| hcodes | H317 - H319 - H412 |
| pcodes | P261 - P273 - P280 - P333 + P313 - P337 + P313 - P362 + P364 |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 2 |
| Flash Point F | does not flash |
| Flash Point C | d
Пуско-наладка
Выполним распаковку, визуальную проверку, сборку и установку, запуск и настройку, проверку функциональности, проведем инструктаж персонала.
Техническое обслуживание
Проведем периодические регламентные работы: визуальный осмотр, очистка засоренных узлов и частей, замена расходных материалов и изношенных деталей, тестирование прибора, проверка показателей и измерение параметров, калибровка и настройка, обновление программного обеспечения.
IQ/OQ/PQ квалификация, валидация
Проведем монтажную (IQ), операционную (OQ) и эксплуатационную (PQ) квалификацию оборудования по протоколам производителя в полном соответствии с стандартами GMP/GLP.
Ремонт
Проведем диагностику в нашем сервисном центре или у Вас на предприятии, выявим причину поломки, устраним неисправность, проведем тестирование, настройку, калибровку отремонтированного оборудования.
Получите коммерческое предложение в течение 1 часаМенеджер подготовит коммерческое предложение и позвонит, если понадобится уточнить детали вашего заказа 
Анна Гловацкая
Менеджер по работе с клиентами
С 2010 года мы поставляем оборудование с заводов Европы. Берем на себя все — от подбора оборудования до внедрения на предприятии
Все сотрудники имеют высшее образование, закончили ведущие химические вузы страны, такие как РХТУ им Менделеева. 
У большинства компаний срок ожидания составляет 10-12 недель. 
Оборудование хранится на сухом отапливаемом складе, где поддерживается ровная температура. 
Работаем с PonyExpress и Деловыми линиями. Вы также можете выбрать свою транспортную компанию или забрать товар со склада в Москве. 
В случае любых неполадок за свой счет выполним ремонт в сервисном центре или на заводе-изготовителе. Или бесплатно заменим прибор на новый. 
Производим пуско-наладку оборудования, валидацию, обучение сотрудников. Если нужно, привлекаем инженеров с заводов- изготовителей. ОтзывыВ числе наших клиентов организации
|
