promega celltiter-fluor cell viability assay protocol

Technical Bulletin

CellTiter-Fluor™ Cell

Viability Assay

INSTRUCTIONS FOR USE OF PRODUCTS G6080, G6081 AND G6082.

PRINTED IN USA.

Revised 12/12Part# TB371

CellTiter-Fluor™ Cell Viability

Assay

All technical literature is available on the Internet at: /protocols/

Please visit the web site to verify that you are using the most current version of this

Technical Bulletin. Please contact Promega Technical Services if you have questions on use

of this system. E-mail: techserv@

1

t Components and .4

t Preparation 5

ols for the CellTiter-Fluor™ Cell 5

A.

B.

C.

D.

E.

Determining Assay Sensitivity, 6

Determining Assay Sensitivity, 7

Example Viability 8

Example Multiplex Assay Protocol (with luminescent caspase assay).......9

10

10

11

12

ption

The CellTiter-Fluor™ Cell Viability Assay

(a)

is a nonlytic, single-reagent-

addition fluorescence assay that measures the relative number of live cells in a

culture population after experimental manipulation (Figures 1 and 2). The

CellTiter-Fluor™ Cell Viability Assay measures a conserved and constitutive

protease activity within live cells and therefore serves as a marker of cell

viability (1). Results obtained using the CellTiter-Fluor™ Cell Viability Assay

correlate well with other established methods of determining cell viability

(Figure 3). The live-cell protease activity is restricted to intact viable cells and is

measured using a fluorogenic, cell-permeant, peptide substrate (glycyl-

phenylalanyl-aminofluorocoumarin; GF-AFC). The substrate enters intact cells

where it is cleaved by the live-cell protease activity to generate a fluorescent

signal proportional to the number of living cells (Figure 4). This live-cell

protease becomes inactive upon loss of cell membrane integrity and leakage

into the surrounding culture medium.

The CellTiter-Fluor™ Cell Viability Assay also can be used in a single-well,

sequential, multiplex format with other downstream chemistries to normalize

data by cell number. Data from the assay can serve as an internal control and

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Printed in USA.

Revised 12/12

Part# TB371

Page 1

allow identification of errors resulting from cell clumping or compound

cytotoxicity. The CellTiter-Fluor™ Cell Viability Assay is compatible with most

Promega luminescence assays or spectrally distinct fluorescence assay methods,

such as assays measuring caspase activation, reporter gene expression or

orthogonal measures of viability. However, some P450-Glo™ multiplex

protocols may require removing culture supernatant to a separate assay well

before performing the assay because of isoform-specific competitive inhibition

of the cytotochrome P450 enzymes by the coumarin product of the CellTiter-

Fluor™ Cell Viability Assay reaction.

Add GF-AFC Substrate

to Assay Buffer to create

the CellTiter-Fluor™ Reagent.

GF-AFC Substrate

Assay Buffer

CellTiter-Fluor™ Reagent

Add reagent to plate in

proportional volumes, mix

and incubate.

Measure

fluorescence.

Figure 1. Schematic diagram of the CellTiter-Fluor™ Cell Viability Assay.

Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA

Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 ·

Part# TB371

Page 2

Printed in USA.

Revised 12/12

6

8

6

8

M

A

Assay Benefits

Measure the Relative Number of Live Cells in Culture: Nonlytic, single-

reagent-addition, homogeneous, “add-mix-measure” protocol.

Get More Data from Every Well: The CellTiter-Fluor™ Cell Viability Assay can

be performed in multiplex with most Promega luminescence assays.

Normalize Data for Cell Number: Normalizing data for live-cell number

makes results more comparable well-to-well, plate-to-plate, day-to-day.

GF

CF

3

Active Live-Cell

LIVE-CELL SUBSTRATE:

cell-permeant fluorogenic

O

substrate for the live-cell

protease (Gly-Phe-AFCoumarin)

Protease

H

2

N

CF

3

GF–N

H

O

O

Live-cell protease

substrate can cross

the cell membrane.

O

Nucleus

Figure 2. CellTiter-Fluor™ Cell Viability Assay chemistry. The cell-permeant substrate

enters the cell, where it is cleaved by the live-cell protease activity to produce the

fluorescent AFC. The live-cell protease is labile in membrane-compromised cells and

cannot cleave the substrate.

A.

A

T

P

-

B

a

s

e

d

A

s

s

a

y

L

u

m

i

n

e

s

c

e

n

c

e

(

R

L

U

)

160,000

120,000

r

2

= 0.9923

80,000

40,000

0

2507501,250

1,7502,2502,750

GF-AFC Fluorescence (RFU)

3,100

3,000

2,900

2,800

2,700

2,600

2,500

5001,0001,100

r

2

= 0.963

B.

R

e

s

o

r

u

f

i

n

F

l

u

o

r

e

s

c

e

n

c

e

(

R

F

U

)

GF-AFC Fluorescence (RFU)

Figure 3. The CellTiter-Fluor™ Cell Viability Assay shows strong correlation with

established methods for measuring viability. Panel A. The GF-AFC Substrate signal

from serial dilutions of live cells plotted against results from the CellTiter-Glo

®

Luminescent Cell Viability Assay (Cat.# G7570), which measures cellular ATP.

Panel GF-AFC Substrate signal from serial dilutions of live cells plotted

against results achieved using the CellTiter-Blue

®

Cell Viability Assay (Cat.# G8080).

Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA

Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 ·

Printed in USA.

Revised 12/12

Part# TB371

Page 3

6

8

6

7

M

A

6

9

9

5

M

A

Live Cell

7,500

A

F

C

F

l

u

o

r

e

s

c

e

n

c

e

(

R

F

U

)

6,500

5,500

4,500

3,500

2,500

1,500

500

0

2,0004,000

6,000

Viable

Treated

8,000

10,000

12,000

6

8

6

6

M

A

Cells or Cell Equivalents/Well

Figure 4. The CellTiter-Fluor™ Cell Viability Assay signal derived from viable

cells (untreated) is proportional to cell number. Dead cells (treated) do not

contribute appreciable signal in the assay.

t Components and Storage Conditions

Size

10ml

Cat.#

G6080

Product

CellTiter-Fluor™ Cell Viability Assay

Cat.# G6080 contains sufficient reagents for 100 assays at 100µl/assay in a 96-well plate

format or 400 assays at 25µl/assay in a 384-well plate format. Includes:

•1 × 10ml

•1 × 10µl

Assay Buffer

GF-AFC Substrate (100mM in DMSO)

Size

5 × 10ml

Cat.#

G6081

Product

CellTiter-Fluor™ Cell Viability Assay

Cat.# G6081 contains sufficient reagents for 500 assays at 100µl/assay in a 96-well plate

format or 2,000 assays at 25µl/well in a 384-well format. Includes:

•5 × 10ml

•5 × 10µl

Assay Buffer

GF-AFC Substrate (100mM in DMSO)

Size

2 × 50ml

Cat.#

G6082

Product

CellTiter-Fluor™ Viability Assay

Cat.# G6082 contains sufficient reagents for 1,000 assays at 100µl/assay in a 96-well plate

format or 4,000 assays at 25µl/well in a 384-well format. Includes:

•2 × 50ml

•2 × 50µl

Assay Buffer

GF-AFC Substrate (100mM in DMSO)

Storage Conditions: Store the CellTiter-Fluor™ Cell Viability Assay

components at –20°C. See product label for expiration date.

Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA

Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 ·

Part# TB371

Page 4

Printed in USA.

Revised 12/12

3.

1.

Reagent Preparation and Storage

Completely thaw the CellTiter-Fluor™ Cell Viability Assay components in a

37°C water bath. Vortex the GF-AFC substrate to ensure homogeneity, then

briefly centrifuge for complete substrate volume recovery.

Transfer the GF-AFC Substrate (10µl for Cat.# G6080 and G6081; 50µl for Cat.#

G6082) into the Assay Buffer container (10ml for Cat.# G6080 and G6081; 50ml

for Cat.# G6082) to form a 2X Reagent. Mix by vortexing the contents until the

substrate is thoroughly dissolved.

Note: The solution may initially appear “milky” when the GF-AFC substrate is

delivered to the buffer. This is normal. The substrate will dissolve with

vortexing. The CellTiter-Fluor™ Reagent may be scaled to accommodate the

volumes required for downstream multiplexes. To do this, use 1/5 the volume

of buffer when you prepare the reagent (i.e., 10µl of the GF-AFC Substrate in

2ml of Assay Buffer). Be sure to label the bottle to indicate that this is a more

concentrated reagent, suitable for multiplex assays. Add the reagent at 1/5 the

volume of the cell culture.

Storage: The CellTiter-Fluor™ Viability Reagent should be used within 24

hours if stored at room temperature. Unused GF-AFC Substrate and Assay

Buffer can be stored at 4°C for up to 7 days with no appreciable loss of

activity.

2.

ols for the CellTiter-Fluor™ Cell Viability Assay

Materials to Be Supplied by the User

•96-, 384-, or 1536-well opaque-walled tissue culture plates compatible with

fluorometer (clear or solid bottom)

•multichannel pipettor or liquid-dispensing robot

•reagent reservoirs

•fluorescence plate reader with filter sets for AFC (380–400nm

Ex

/505

Em

)

•orbital plate shaker

•compound known to cause 100% cytotoxicity or lytic detergent (digitonin,

Calbiochem Cat.# 300410 or Sigma-Aldrich Cat.# D141 at 20mg/ml in DMSO).

If you have not performed this assay on your cell line previously, we

recommend determining assay sensitivity using your cells and one of the two

methods described below (Section 4.A or 4.B). If you do not need to determine

assay sensitivity for your cells, proceed to Section 4.C.

Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA

Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 ·

Printed in USA.

Revised 12/12

Part# TB371

Page 5

ining Assay Sensitivity, Method 1

t adherent cells (by trypsinization, etc.), wash with fresh medium (to

remove residual trypsin) and resuspend in fresh medium.

Note:For cells growing in suspension, proceed to Step 2.

ine the number of viable cells by trypan blue exclusion using a

hemacytometer, then adjust them by dilution to 100,000 viable cells/ml in

at least 3.0ml of fresh medium.

Note: Concentrate the cells by centrifuging and removing medium if the

cell suspension is less than 100,000 cells/ml.

100µl of the 100,000 cell/ml dilution (10,000 cells/well) to all wells of

rows A and B in a 96-well plate (Table 1).

100µl of fresh medium to all wells in rows B–H.

a multichannel pipettor, mix the cell suspension in row B by

pipetting (being careful not to create foaming or bubbles). Transfer 100µl

from row B to row C. Repeat mixing and transfer 100µl from row C to row

D. Continue this process to row G. After mixing the diluted suspension at

row G, aspirate 100µl from wells and discard it. This procedure dilutes

your cells from 10,000 cell/well in row A to 156 cells/well in row G. Row

H will serve as the no-cell, background control.

Table 1. Schematic of 96-Well Plate Layout.

1

A

B

C

D

E

F

G

H

23456789101112

10,000 cells/well

5,000 cells/well

2,500 cells/well

1,250 cells/well

625 cells/well

313 cells/well

156 cells/well

0 cells/well

digitonin to 300µg/ml in water. Using a multichannel pipet,

carefully add 10µl of the diluted digitonin to all wells of columns 7–12 to

lyse cells (treated samples). Add 10µl of water to all wells of columns 1–6

to normalize the volume (untreated cells).

100µl of the CellTiter-Fluor™ Reagent to all wells, mix briefly by

orbital shaking and incubate at 37°C for at least 30 minutes.

Note:Longer incubations may improve assay sensitivity and dynamic

range. However, do not incubate longer than 3 hours, and be sure to shield

plates from ambient light.

Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA

Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 ·

Part# TB371

Page 6

Printed in USA.

Revised 12/12

e resulting fluorescence with a fluorometer (380–400nm

Ex

/505nm

Em

)

Note: You may need to adjust instrument gains (applied photomultiplier

tube energy).

ate the practical sensitivity for your cell type by making a signal-to-

noise calculation for each dilution of cells (10,000 cells/well; 5,000

cells/well; 2,500 cells/well, etc.).

Viability S:N =

(Average Untreated – Average Treated)

Std. Dev. of H-1 through H-6

Note: The practical level of assay sensitivity for the assay is a signal-to-

noise ratio of greater than 3 standard deviations (derived from reference 1).

ining Assay Sensitivity, Method 2

t adherent cells (by trypsinization, etc.), wash with fresh medium

(to remove residual trypsin) and resuspend in fresh medium.

Note:For cells growing in suspension, proceed to Step 2.

ine the number of viable cells by trypan blue exclusion using a

hemacytometer, then adjust the cells by dilution to 100,000 viable cells/ml

in at least 20ml of fresh medium.

Note:Concentrate the cells by centrifuging and removing medium if the

pool of cells is less than 100,000 cells/ml.

the volume of diluted cells into separate tubes. Subject one tube to

"moderate" sonication (empirically determined by postsonication

morphological examination) to rupture cell membrane integrity and to

simulate a 100% dead population. The second tube of untreated cells will

serve as the maximum viable population.

a spectrum of viability by blending sonicated and untreated

populations in 1.5ml microcentrifuge tubes as described in Table 2.

Table 2. Spectrum of Viability Generated by Blending Sonicated and Untreated Cells.

Percent Viability

100

95

90

75

50

25

10

5

0

µl Sonicated

0

50

100

250

500

750

900

950

1,000

µl Untreated

1,000

950

900

750

500

250

100

50

0

Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA

Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 ·

Printed in USA.

Revised 12/12

Part# TB371

Page 7

ining Assay Sensitivity, Method 2 (continued)

mixing each blend by gently vortexing, pipet 100µl of each blend into

8 replicate wells of a 96-well plate. Add the 100% viable cells to column 1,

95% viable to column 2, etc. Add cell culture medium only to column 10 to

serve as a no-cell control.

CellTiter-Fluor™ Reagent in an equal volume (100µl per well) to all

wells, mix briefly by orbital shaking, then incubate for at least 30 minutes

at 37°C.

Note:Longer incubations may improve assay sensitivity and dynamic

range. However, do not incubate longer than 3 hours, and be sure to shield

plates from ambient light.

e resulting fluorescence with a fluorometer (380–400nm

Ex

/505nm

Em

).

Note: You may need to adjust instrument gains (applied photomultiplier

tube energy).

ate the practical sensitivity for your cell type by making a signal-to-

noise calculation for each blend of cell viability (X = 95, 90%, etc.).

(Average 100% – Average X%)

Viability S:N =

Standard Deviation of 0% (viable cells)

Note: The practical level of assay sensitivity for the assay is a signal-to-

noise ratio of greater than 3 standard deviations (derived from reference 1).

e Viability Assay Protocol

up 96-well assay plates containing cells in culture medium at desired

density.

test compounds and vehicle controls to appropriate wells so that the

final volume is 100µl in each well (25µl for a 384-well plate).

e cells for the desired test exposure period.

CellTiter-Fluor™ Reagent in an equal volume (100µl per well) to all

wells, mix briefly by orbital shaking, then incubate for at least 30 minutes

at 37°C.

Note:Longer incubations may improve assay sensitivity and dynamic

range. However, do not incubate more than 3 hours, and be sure to shield

plates from ambient light.

e resulting fluorescence using a fluorometer (380–400nm

Ex

/505nm

Em

).

Note:You may need to adjust instrument gains (applied photomultiplier

tube energy).

Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA

Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 ·

Part# TB371

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Printed in USA.

Revised 12/12

e Multiplex Assay Protocol (with luminescent caspase assay)

up 96-well assay plates containing cells in culture medium at the

desired density.

test compounds and vehicle controls to appropriate wells so that the

final volume is 100µl in each well (25µl for a 384-well plate).

e cells for the desired test exposure period.

Note:Caspase activation is a transient event dictated by compound

potency and cell cycle susceptibility. Time course experiments are often

useful for defining peak caspase activity and cytotoxicity.

20µl of CellTiter-Fluor™ Reagent (prepared as 10µl substrate in 2ml

Assay Buffer) to all wells, and mix briefly by orbital shaking. Incubate for

at least 30 minutes at 37°C.

Note: Longer incubations may improve assay sensitivity and dynamic

range. However, do not incubate longer than 3 hours, and be sure to shield

plates from ambient light.

e resulting fluorescence using a fluorometer (380–400nm

Ex

/505nm

Em

).

Note: You may need to adjust instrument gains (applied photomultiplier

tube energy).

an equal volume of Caspase-Glo

®

3/7 Reagent prepared as described

in Technical Bulletin #TB323 to wells (100–120µl per well), incubate for 30

minutes, then measure luminescence using a luminometer.

CellTiter-Fluor™ Cell Viability Assay

Caspase-Glo

®

3/7 Assay

6,000

30,000

L

u

m

i

n

e

s

c

e

n

c

e

(

R

L

U

)

4,000

20,000

10,000

2,000

0

–11

0

–10–9–8–7

6

8

6

5

M

A

log

10

[paclitaxel] M

Figure 5. Multiplex of CellTiter-Fluor™ Assay and Caspase-Glo

®

3/7

CellTiter-Fluor™ Reagent was added to wells and viability measured after

incubation for 30 minutes at 37°C. Caspase-Glo

®

3/7 Reagent was added and

luminescence measured after a 30-minute incubation (10,000 cells/well).

Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA

Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 ·

Printed in USA.

Revised 12/12

Part# TB371

Page 9

F

l

u

o

r

e

s

c

e

n

c

e

(

R

F

U

)

ended Controls

No-Cell Control: Set up triplicate wells without cells to serve as a control to

determine background fluorescence.

Untreated Cells Control:Set up triplicate wells with untreated cells to serve as

a vehicle control. Add the same solvent used to deliver the test compounds to

the vehicle control wells.

Optional Test Compound Control: Set up triplicate wells without cells but

containing the vehicle and test compound to test for possible interference with

the assay chemistry.

Positive Control for Viability:Set up triplicate wells containing cells treated

with a compound known to be toxic to the cells used in your model system.

l Considerations

Optical Filters and Instrumentation:Fluorogenic dyes exhibit distinct

absorption (excitation) and emission profiles when a light energy source is

applied. Most fluorometers or multimode instruments contain optical band-

pass filters that restrict the wavelengths of light used to excite a fluorophore

and the wavelengths passing through to the detector. Note that deviation from

the optimal filter set recommendations (Figure 6) may adversely affect assay

sensitivity and performance.

3,500

3,000

Excitation

Emission

F

l

u

o

r

e

s

c

e

n

c

e

(

R

F

U

)

2,500

2,000

1,500

1,000

500

0

250

Wavelength (nm)

Figure 6. Optimal excitation and emission spectra for AFC.

Background Fluorescence and Inherent Serum Activity:Tissue culture

medium that is supplemented with animal serum may contain detectable levels

of the protease marker used to measure live-cells. This protease activity may

vary among different lots of serum. To correct for variability, determine

background fluorescence using samples containing medium plus serum

without cells.

Temperature: The generation of fluorescent product is proportional to the live-

cell protease activity. The activity of this protease is influenced by temperature.

Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA

Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 ·

Part# TB371

Page 10

Printed in USA.

Revised 12/12

6

8

6

4

M

A

For best results, we recommend incubating at a constant controlled temperature

to ensure uniformity across the plate. After adding reagent and briefly mixing,

we suggest one of two options:

37°C in a water-jacketed incubation module (Me’Cour, etc.).

Note:Incubation at 37°C in a CO

2

culture cabinet may lead to edge-effects

resulting from thermal gradients.

room temperature with or without orbital shaking.

Note: Assays performed at room temperature may require more than

30minutes of incubation for optimal sensitivity. However, do not incubate

longer than 3 hours.

Assay Controls: In addition to a no-cell control to establish background

fluorescence, we recommend including a maximum viability (untreated cells)

and maximum cytotoxicity control in the experimental design. The maximum

viability control is established by adding vehicle only (used to deliver the test

compound to test wells). In most cases, this consists of a buffer system or

medium and the equivalent amount of solvent added with the test compound.

The maximum cytotoxicity control can be determined using a compound that

causes 100% cytotoxicity or a lytic compound added to compromise viability

(digitonin). See Section 4.A.

Viability Marker Half-Life:The activity of the protease marker found has no

half-life in viable cells. Viable cells will process the substrate to liberate the AFC

fluorophore. However, when cells lose membrane integrity, the protease activity

declines very quickly. Therefore enzymatic instability of the live-cell protease

outside of viable cells establishes GF-AFC as a good marker for cell viability.

Light Sensitivity: Although the GF-AFC Substrate demonstrates good general

photostability, the liberated AFC fluorophore (after contact with protease) can

degrade with prolonged exposure to ambient light sources. We recommend

shielding the plates from ambient light at all times.

Cell Culture Medium:The GF-AFC Substrate is introduced into the test well

using an optimized buffer system that mitigates differences in pH from

treatment. In addition, the buffer system supports protease activity in a host of

different culture media with varying osmolarity. With the exception of media

formulations with either very high serum content or phenol red indicator, no

substantial performance differences will be observed among media.

nces

, A.L. et al. (2007) A homogeneous assay to measure live and dead cells in the

same sample by detecting different protease markers. Anal. Biochem.366, 197–206.

, J-H. et al.(1999) A simple statistical parameter for use in evaluation and

validation of high-throughput screening assays. J. Biomol. Screen. 4, 67–73.

Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA

Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 ·

Printed in USA.

Revised 12/12

Part# TB371

Page 11

d Products

Cell Viability and CytotoxicityAssays

Product

MultiTox-Fluor Multiplex Cytotoxicity Assay

MultiTox-Glo Multiplex Cytotoxicity Assay

CytoTox-Glo™ Cytotoxicity Assay

CytoTox-Fluor™ Cytotoxicity Assay

CellTiter-Glo

®

Luminescent Cell Viability Assay

CytoTox-ONE™ Homogeneous Membrane

Integrity Assay

CellTiter-Blue

®

Cell Viability Assay

Additional Sizes Available.

Size

10ml

10ml

10ml

10ml

10ml

1,000–4,000 assays

20ml

Cat.#

G9200

G9270

G9290

G9260

G7570

G7891

G8080

Apoptosis Assays

Product

Caspase-Glo

®

2 Assay

Caspase-Glo

®

6 Assay

Caspase-Glo

®

3/7 Assay

Caspase-Glo

®

8 Assay

Caspase-Glo

®

9 Assay

Apo-ONE

®

Homogeneous Caspase 3/7 Assay

Additional Sizes Available.

Size

10ml

10ml

10ml

10ml

10ml

10ml

Cat.#

G0940

G0970

G8091

G8201

G8211

G7790

Reporter Gene Assays

Product

Bright-Glo™ Luciferase Assay System

Steady-Glo

®

Luciferase Assay System

Additional Sizes Available.

Size

10ml

10ml

Cat.#

E2610

E2510

(a)

Patent Pending.

© 2007–2012 Promega Corporation. All Rights Reserved.

Apo-ONE, Caspase-Glo, CellTiter-Blue, CellTiter-Glo and Steady Glo are registered trademarks of Promega Corporation.

BrightGlo, CellTiter-Fluor, CytoTox-Fluor, CytoTox-Glo, CytoTox-ONE and P450-Glo and are trademarks of Promega

Corporation.

Products may be covered by pending or issued patents or may have certain limitations. Please visit our Web site for more

information.

All prices and specifications are subject to change without prior notice.

Product claims are subject to change. Please contact Promega Technical Services or access the Promega online catalog for the

most up-to-date information on Promega products.

Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA

Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 ·

Part# TB371

Page 12

Printed in USA.

Revised 12/12