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Merck

04511

Supelco

Live/Dead Cell Double Staining Kit

suitable for fluorescence

Sinónimos:

Staining kit for live/dead cells

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About This Item

Código UNSPSC:
12161503
NACRES:
NA.32
En este momento no podemos mostrarle ni los precios ni la disponibilidad

idoneidad

suitable for fluorescence

Nivel de calidad

temp. de almacenamiento

−20°C

Aplicación

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.

Solo componentes del kit

Referencia del producto
Descripción

  • Solution A (Calcein AM solution) 4 × 50

  • Solution B (propidium iodide solution) 300 μL

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Código de clase de almacenamiento

10 - Combustible liquids

Clase de riesgo para el agua (WGK)

WGK 2

Punto de inflamabilidad (°F)

185.0 °F - closed cup

Punto de inflamabilidad (°C)

85 °C - closed cup


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T Matsuse et al.
Journal of clinical pathology, 51(7), 515-519 (1998-11-03)
To investigate the presence and distribution of advanced glycation end products (AGE) in pulmonary fibrosis. Lung tissue samples obtained from seven necropsy cases with idiopathic pulmonary fibrosis and seven with normal pulmonary parenchyma were examined immunohistochemically with a monoclonal antibody
T Kimura et al.
Neuroscience letters, 208(1), 53-56 (1996-04-12)
The recent immunological demonstration of advanced glycation end products (AGE) of the Maillard reaction in several human tissues suggests a possible involvement of AGE in the aging process. We previously prepared a monoclonal anti-AGE antibody (6D12) which recognized N epsilon-(carboxymethyl)lysine.
Amin Hassanzadeh-Barforoushi et al.
Lab on a chip, 18(15), 2156-2166 (2018-06-21)
We present here a new method to easily and reliably generate an array of hundreds of dispersed nanoliter-volume semi-droplets for single-cells culture and analysis. The liquid segmentation step occurs directly in indexed traps by a tweezer-like mechanism and is stabilized
Camille Raillon et al.
Cytometry. Part A : the journal of the International Society for Analytical Cytology, 95(10), 1085-1095 (2019-08-01)
The isolation, analysis, and enumeration of circulating tumor cells (CTCs) from cancer patient blood samples are a paradigm shift for cancer patient diagnosis, prognosis, and treatment monitoring. Most methods used to isolate and enumerate these target cells rely on the
S Yoshida et al.
Clinical nephrology, 49(5), 273-280 (1998-06-09)
Cardiovascular disease is one of the most common complications of dialysis and renal transplant patients, and high levels of AGE are present in end-stage renal failure. To address the potential involvement of AGE and growth factors in the pathophysiology of

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Questions

1–3 of 3 Questions  
  1. Can this kit be used to assess cell viability within bacterial biofilms?

    1 answer
    1. 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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  2. Can the live/dead double staining kit be used on fixed cells?

    1 answer
    1. 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?

  3. How many assays or tests can be conducted using one kit of PN: 04511-1KT-F?

    1 answer
    1. 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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