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MAN0018370 PrestoBlueCellViabilityReagent PI

MAN0018370-PrestoBlueCellViabilityReagent-PI

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30 views

MAN0018370 PrestoBlueCellViabilityReagent PI

MAN0018370-PrestoBlueCellViabilityReagent-PI

Uploaded by

calidad ama
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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PRODUCT INFORMATION SHEET

PrestoBlue™ Cell Viability Reagent


Catalog Numbers A13261 and A13262
Pub. No. MAN0018370 Rev. B.0
WARNING! Read the Safety Data Sheets (SDSs) and follow the handling instructions. Wear appropriate protective eyewear,
clothing, and gloves. Safety Data Sheets (SDSs) are available from thermofisher.com/support.

Contents and storage


Sufficient material is supplied for 1,000 × 96-well assays (Cat. No. A13261) or 10,000 × 96-well assays (Cat. No. A13262) based on the
protocol that is described in this document.

Component Cat. No. A13261 Cat. No. A13262 Storage[1]


• Store at 2–8°C.
PrestoBlue™ Cell Viability Reagent 1 × 25 mL (10X) 1 × 100 mL (10X)
• Protect from light.
[1] When stored as directed, the kit is stable for 6 months from date of receipt.

• Fluorescence excitation and emission ranges: 540–570 nm and 580–610 nm.


Fluorescence excitation/emission maxima: 560/590 nm
• Absorbance: Monitor the absorbance at 570 nm, using 600 nm as a reference wavelength (normalized to the 600-nm value).
™
The PrestoBlue Cell Viability Reagent is a complete add and read,
Product description nontoxic reagent that does not require cell lysis. Since no lysis is
™
Measuring changes in cell viability is a fundamental method for required the diluted PrestoBlue solution can be removed, then
evaluating cell health, determining genotoxicity, and evaluating replaced with complete growth media and the cells cultured
anti-cancer drugs. Cell health can be monitored by detecting further.
changes in several key indicators. These include changes to plasma ™
Based on the cell type and incubation time, PrestoBlue Cell
membrane integrity, DNA synthesis, DNA content, enzyme Viability Reagent can detect as few as 12 cells per well in a
activity, presence of ATP, and cellular reducing environment. 384-well plate. After ten minutes of incubation, the PrestoBlue
™

Monitoring changes to the cellular reducing environment or reagent can linearly detect cells down to 98 cells/well with a Z'
metabolic activity by using resazurin-based reagents is a well- value of greater than 0.5.
established and reliable indicator of cell viability or death.
™
Resazurin, the active ingredient of PrestoBlue Cell Viability
Reagent, is a nontoxic, cell-permeable compound that is blue in
Seed and treat cells
color and virtually non-fluorescent. On entering live cells, the 1. One day before your experiment, seed cells into a 96-well
cellular reducing environment reduces resazurin to resorufin, a plate containing 100 µL/well of cell culture medium.
compound that is red and highly fluorescent. Viable cells
Note: If you are using a 384-well plate, use 50 µL/well of cell
continuously convert resazurin to resorufin, increasing the overall culture medium.
fluorescence and color of the media surrounding the cells. Also,
the conversion of resazurin to resorufin results in a pronounced 2. Incubate the cells overnight in a 37°C incubator using the
color change, therefore cell viability can be detected using appropriate CO2 percentage.
absorbance-based plate readers.
™ 3. (Optional) Treat cells with the test compound 24–72 hours
Unlike other resazurin-based reagents, PrestoBlue has been before performing the viability with the cell viability reagent.
formulated with a proprietary buffering system that resulted in a
reagent with a physiological pH range optimal for the fast
determination of mammalian cell viability. Incubation times using
™
PrestoBlue are shorter than with other resazurin-based cell
viability reagents. Cell viability can be detected with a short 10-
™
minute incubation with the PrestoBlue Cell Viability Reagent.

For Research Use Only. Not for use in diagnostic procedures.


4. Record results using the following fluorescence or
Determine cell viability absorbance values:

1. Warm the cell viability reagent to room temperature before • Fluorescence: Read fluorescence using a fluorescence
use. excitation wavelength of 560 nm (excitation range is
540–570 nm) and an emission of 590 nm (emission range
is 580–610–nm).
2. Add 1/10th volume of cell viability reagent directly to cells in
culture medium according to the following table. • Absorbance: Monitor the absorbance of reagent at
570 nm, using 600 nm as a reference wavelength
Volume of 10X cell (normalized to the 600-nm value).
Volume of cells +
Format viability reagent to
medium Note: Assay plates or tubes can be wrapped in foil, stored at
add
4°C, then read in 1–3 days without affecting the fluorescence
Cuvette 900 µL 100 µL or absorbance values.
96-well plate 90 µL 10 µL
5. (Optional) Add 50 µL of 3% SDS directly to 100 µL of cells in
384-well plate 36 µL 4 µL cell viability reagent to stop the reaction.

3. Incubate for ≥10 minutes at 37°C in a cell culture


incubator, protected from direct light.
Limited product warranty
Life Technologies Corporation and/or its affiliate(s) warrant their
Note: Longer incubation times increase the sensitivity of
products as set forth in the Life Technologies' General Terms and
detection. We recommend using up to a 3-hour incubation
Conditions of Sale at www.thermofisher.com/us/en/home/global/
time to increase sensitivity. As this is a live cell assay,
terms-and-conditions.html. If you have any questions, please
readings can be taken at multiple time points.
contact Life Technologies at www.thermofisher.com/support.

Life Technologies Corporation | 29851 Willow Creek | Eugene, OR 97402


For descriptions of symbols on product labels or product documents, go to thermofisher.com/symbols-definition.

The information in this guide is subject to change without notice.


DISCLAIMER: TO THE EXTENT ALLOWED BY LAW, THERMO FISHER SCIENTIFIC INC. AND/OR ITS AFFILIATE(S) WILL NOT BE LIABLE FOR SPECIAL, INCIDENTAL, INDIRECT,
PUNITIVE, MULTIPLE, OR CONSEQUENTIAL DAMAGES IN CONNECTION WITH OR ARISING FROM THIS DOCUMENT, INCLUDING YOUR USE OF IT.
Important Licensing Information: These products may be covered by one or more Limited Use Label Licenses. By use of these products, you accept the terms and conditions of all
applicable Limited Use Label Licenses.
©2019 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified.

thermofisher.com/support | thermofisher.com/askaquestion
thermofisher.com
7 January 2019

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