How does the Alamar blue assay work?

How does the Alamar blue assay work?

AlamarBlue Cell Viability Reagent quantitatively measures the proliferation of mammalian cell lines, bacteria and fungi. The dye incorporates an oxidation-reduction (REDOX) indicator that both fluoresces and change color in response to the chemical reduction due to cell growth.

How is Alamar blue calculated?

O1 = molar extinction coefficient (E) of oxidized alamarBlue (blue) at 570 nm* O2 = E of oxidized alamarBlue at 600 nm* A1 = absorbance of test wells at 570 nm. A2 = absorbance of test wells at 600 nm….62%

What is microplate Alamar blue assay?

The 96-well Microplate Alamar Blue Assay (MABA) allows for the quantitative determination of drug susceptibility against any strain of replicating Mycobacterium tuberculosis to be completed within a week at minimal cost. tuberculosis is an important factor when developing drug candidates against M. tuberculosis.

What is Alamar?

Alamar Blue Cell Viability Assay (6 Citations) AlamarBlue Cell Viability Assay reagent quantitatively measures the proliferation of mammalian cell lines, bacteria and fungi. In alamarBlue assay the growing cells cause a chemical reduction of the alamarBlue dye from non-fluorescent blue to red fluorescent.

How do you make Alamar blue solution?

Reagent Preparation

1. Dissolve alamar blue in PBS (pH 7.4) to 0.15 mg/ml.
2. Filter-sterilize the resazurin solution through a 0.2 μm filter into a sterile, light protected container.
3. Store the resazurin solution protected from light at 4°C for frequent use or at -20°C for long term storage.

What is Presto blue?

PrestoBlue® is a ready to use cell permeable resazurin-based solution that functions as a cell viability indicator by using the reducing power of living cells to quantitatively measure the proliferation of cells. This color change can be detected using fluorescence or absorbance measurements.

Is Alamar blue light sensitive?

alamarBlue® should be stored in the dark, since the compound is light sensitive (Table 2). The product may be stored for 12 months at room temperature.

How does resazurin reduction assay work?

The resazurin assay protocol is based on the reduction of oxidized non-fluorescent blue resazurin to a red fluorescent dye (resorufin) by the mitochondrial respiratory chain in live cells. The amount of resorufin produced is directly proportional to the number of living cells.

What does the Alamar Blue cell viability assay measure?

Alamar Blue Cell Viability Assay. AlamarBlue Cell Viability Assay reagent quantitatively measures the proliferation of mammalian cell lines, bacteria and fungi. The dye incorporates an oxidation-reduction (REDOX) indicator that both fluoresces and change color in response to the chemical reduction of growth medium due to cell growth.

What is the oxidation state of Alamar Blue?

The oxidation-reduction potential of Alamar Blue is +380 mV at pH 7.0, 25 °C. Alamar Blue, therefore, can be reduced by NADPH (E o = 320 mV), FADH (E o = 220 mV), FMNH (E o = 210 mV), NADH (E o = 320 mV), as well as the cytochromes (E o = 290 mV to +80 mV).

What is alamarblue reagent used for?

AlamarBlue Cell Viability Assay reagent quantitatively measures the proliferation of mammalian cell lines, bacteria and fungi. The dye incorporates an oxidation-reduction (REDOX) indicator that both fluoresces and change color in response to the chemical reduction of growth medium due to cell growth.

What are the precautions for Alamar Blue assays?

Assays must be carried out at a uniform temperature to ensure reproducibility of all wells and across a single plate. Alamar Blue is photosensitive and the incubations must be done in the dark. The culture medium and the test compound themselves should not interact with the assay chemistry.

How do you assay cell proliferation?

Cell proliferation may be studied by monitoring the incorporation of a radioisotope, [3H]-thymidine, into cellular DNA, followed by autoradiography. Alternatively, 5-bromo-2′-deoxy-uridine (BrdU assays) may be used instead of thymidine.

What do proliferation assays detect?

Cell proliferation assays typically detect changes in the number of cells in a division or changes in a cell population. Cell proliferation assays are mainly divided into four methods: metabolic activity assays, cell proliferation marker assays, ATP concentration assays, and DNA synthesis assays.

What are cell proliferation markers?

For example, Ki67, proliferating cell nuclear antigen (PCNA), and minichromosome maintenance (MCM) proteins are standard markers of proliferation that are commonly used to assess the growth fraction of a cell population. The function of Ki67, the widely used marker of proliferation, still remains unclear.

What is the Alamar Blue bioassay?

The Alamar Blue bioassay has been utilized over the past 50 years to assess cell viability and cytotoxicity in a range of biological and environmental systems and in a number of cell types including bacteria, yeast, fungi, protozoa and cultured mammalian and piscine cells.

Why use alamarblue to evaluate cell viability?

Analysis of cell proliferation and cytotoxicity is a vital step in evaluating cellular health and in the drug discovery process. alamarBlue is a proven cell viability indicator that uses the natural reducing power of living cells to convert resazurin to the fluorescent molecule, resorufin.

alamarBlue® 1 Cell proliferation assays. 2 The reagent can be used to establish proliferation or relative cytotoxicity in a cell proliferation assay. 3 Baseline data for predicting the toxicity of related novel agents can be compared to baseline data with known in-vivo toxicity.

What is the optimum incubation time for alamarblue?

Incubate cultures with alamarBlue for 4-8 hr. N.B. The optimum incubation time may vary between cell types. Measure cytotoxicity or proliferation using spectrophotometry or fluorescence. Harvest cells which are in the log phase of growth and determine cell count. Adjust the cell count to 1 x 10 4 cells/ml (suggested cell density).