This kit is designed for use with eukaryotic cells and has not been validated for use with bacterial cells.
04511
Doppelfärbungs-Kit für lebende/tote Zellen
suitable for fluorescence
Synonym(e):
Färbungskit für lebende/tote Zellen
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NACRES:
NA.32
UNSPSC Code:
12161503
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Dieser Artikel | |||
|---|---|---|---|
| suitability suitable for fluorescence | suitability - | suitability - | suitability - |
| storage temp. −20°C | storage temp. - | storage temp. - | storage temp. - |
| Quality Level 100 | Quality Level 200 | Quality Level 100 | Quality Level 100 |
Application
Das Lebende/Tote Zellen-Doppelfärbungskit wird für den gleichzeitigen Fluoreszenznachweis von lebenden und toten Zellen verwendet. Dieses Kit enthält Calcein-AM- und Propidiumiodid(PI)-Lösungen, die lebende bzw. tote Zellen färben. Calcein-AM, das Acetoxymethylester von Calcein, ist stark lipophil mit einer durchlässigen Zellmembran. Obwohl Calcein-AM selbst kein fluoreszierendes Molekül ist, strahlt das Calcein, das in einer lebenden Zelle durch Esterase aus Calcein-AM gewonnen wird, eine starke, grüne Fluoreszenz aus (λex 490 nm, λem 515 nm). Daher färbt Calcein-AM nur lebende Zellen. Alternativ dazu kann der den Kern färbende Farbstoff PA die Membran lebender Zellen nicht passieren. Er erreicht den Kern, indem er ungeordnete Bereiche der Membran toter Zellen passiert und sich in der DNA-Doppelhelix der Zelle einlagert, um eine rote Fluoreszenz auszustrahlen (λex 535 nm, λem 617 nm). Da sowohl Calcein als auch PI-DNA mit einem 490-nm-Licht angeregt werden können, ist die gleichzeitige Überwachung von lebenden und toten Zellen mit einem Fluoreszenzmikroskop möglich. Bei Verwendung von λex 545 nm können nur tote Zellen beobachtet werden.
Nur Kit-Komponenten
Produkt-Nr.
Beschreibung
- Solution A (Calcein AM solution) 4 × 50
- Solution B (propidium iodide solution) 300 μL
Ähnliches Produkt
Produkt-Nr.
Beschreibung
Preisangaben
Lagerklasse
10 - Combustible liquids
wgk
WGK 2
flash_point_f
185.0 °F - closed cup
flash_point_c
85 °C - closed cup
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Verwandter Inhalt
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This kit can be used in bacteria or is just for eukaryotic cells?
1 answer-
Helpful?
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I want to use this kit for suspension cells. Protocol says to prepare a cell suspension with PBS at 1-10 x 10^5 cells/mL and then add 100uL of assay solution. Is it 100uL per mL of cell suspension?. Also, what is the concent of the original solutions?
1 answer-
200 microliters of cell suspension is mixed with 100 microliters of assay solution, and the mixture is incubated at 37 ºC.
Lot-specific concentrations of Solutions A and B are listed in the product's Certificate of Analysis.Helpful?
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I want to use this kit for suspension cells. Protocol says I first need to prepare a cell suspension with PBS at 1-10 x 10^5 cells/mL and then add 100uL of assay solution. Is it 100uL per mL of cell suspension?. What is the conc of the original solutions?
1 answer-
The original manufacturer of this kit recommends adding 100 microliters of assay solution to 200 microliters of the cell suspension. Unfortunately, this key bit of information is missing from the protocol. Each vial of Solution A contains a 1 mM solution of Calcein-AM in DMSO. The vial of Solution B contains a 1.5 mM solution of Propidium Iodide in pure water.
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Can this kit be used to assess cell viability within bacterial biofilms?
1 answer-
This product requires preparation of a cell suspension and has not been validated for bacterial biofilms or cells attached to any surface.
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Can the live/dead double staining kit be used on fixed cells?
1 answer-
The kit is not designed for use with just dead cells. When cells are fixed, it typically results in cell death for all cells. Consequently, if all the cells are dead, it is expected that the Calcein AM would not be able to enter the cells, as Calcein AM is not a fluorescent molecule. The available data sheet states, "While the cells are viable, the calcein generated from CalceinAM by esterase in a viable cell emits strong green fluorescence (excitation: 490 nm, emission: 515 nm)." Therefore, Calcein-AM only stains viable cells. If all the cells are dead, the only observed staining will be from the Propidium Iodide - which is the dead cell stain reagent in kit 04511.
If the cells are stained while viable, it is possible to image the cells after they have been fixed in paraformaldehyde or other fixatives. Although this is a common practice in some laboratories, It's suggested to consider a 10-minute fixation instead of 30 minutes, as fixing for 30 minutes may not be advisable.Helpful?
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How many assays or tests can be conducted using one kit of PN: 04511-1KT-F?
1 answer-
The 04511 kit consists of 2 components, with reagent B being the limiting reagent, requiring just 300 microliters of reagent. To prepare the staining solution, 10 microliters of Reagent A and 5 microliters of Reagent B are mixed with 5 ml of PBS. Each test necessitates 100 microliters of the staining solution. With 5 microliters of Reagent B being sufficient for 50 tests, 300 microliters of Reagent B is adequate for 60 batches of the staining reagent, resulting in 3000 tests (50 x 60). Although the maximum number of specimens might not always be stained in practice, the kit theoretically should allow for 3000 tests.
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