This kit is designed for use with eukaryotic cells and has not been validated for use with bacterial cells.
04511
Live/Dead Cell Double Staining Kit
suitable for fluorescence
Synonyme(s) :
Staining kit for live/dead cells
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| Conditionnement | Référence | Disponibilité | Prix |
|---|---|---|---|
| 1 kit | Vérifier la disponibilité des articles du panier | 1 490,00 $ |
A propos de cet article
NACRES:
NA.32
UNSPSC Code:
12161503
Service technique
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The Live/Dead Cell Double Staining Kit is utilized for simultaneous fluorescence staining of viable and dead cells. This kit contains calcein-AM and propidium iodide (PI) solutions, which stain viable and dead cells, respectively. Calcein-AM, acetoxymethyl ester of calcein, is highly lipophilic and cell membrane permeable. Though calcein-AM itself is not a fluorescent molecule, the calcein generated from Calcein-AM by esterase in a viable cell emits a strong green fluorescence (λex 490 nm, λem 515 nm). Therefore, calcein-AM only stains viable cells. Alternatively, the nuclei staining dye PI cannot pass through a viable cell membrane. It reaches the nucleus by passing through disordered areas of dead cell membrane, and intercalates with the DNA double helix of the cell to emit red fluorescence (λex 535 nm, λem 617 nm). Since both calcein and PI-DNA can be excited with 490 nm light, simultaneous monitoring of viable and dead cells is possible with a fluorescence microscope. Using λex 545 nm, only dead cells can be observed.
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| Quality Level 100 | Quality Level 200 | Quality Level 100 | Quality Level 100 |
| storage temp. −20°C | storage temp. - | storage temp. - | storage temp. 2-8°C |
| suitability suitable for fluorescence | suitability - | suitability - | suitability - |
Composants de kit seuls
Réf. du produit
Description
- Solution A (Calcein AM solution) 4 × 50
- Solution B (propidium iodide solution) 300 μL
Classe de stockage
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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| Référence | GTIN |
|---|---|
| 04511-1KT-F | 04061835183869 |
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This kit can be used in bacteria or is just for eukaryotic cells?
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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?
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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?
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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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How many assays or tests can be conducted using one kit of PN: 04511-1KT-F?
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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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Can the live/dead double staining kit be used on fixed cells?
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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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