THE NEWSLETTER OF ACTIVE MOTIF — October 2007 • volume 8 • number 4

you’ve got to look inside
to learn about regulation,
chromatin & histones

IN
THIS
ISSUE

2 NEW: DNA Damage Assay – an Easy Method
to Assay for Apoptosis

5 Create Chromatin In Vitro to Reveal
Regulatory Mechanisms

3 Simpliﬁed Study of SUMOylation Effects
with SUMOlink™

6 Improved ChIP-IT™ Express Greatly
Reduces Background

3 Easily Compare CpG Methylation ofYour Samples

7 Simpliﬁed Co-Immunoprecipitation of
Intact Nuclear Complexes

4 NEW: Isolate Pure, Separate Fractions ofHistone H2A/H2B and H3/H4 While
Preserving Post-translational Modiﬁcations

8 Learn More Faster With
AlmaKnowledgeServer 2

H2AX DNA Damage

NEW: DNA Damage Assay – an Easy Method to Assay for Apoptosis
Active Motif’s DNA Damage Assay is a simple and fast 2-color ﬂuorescentassay of H2AX phosphorylation, which occurs following double-stranded
breaks in DNA. This makes it a sensitive and accurate assay of apoptosis.

120
100

What’s in the kit?
The DNA Damage Assay includes thepolyclonal phospho-Histone H2AX (Ser139) antibody, Chromeo 488 Goat antiRabbit secondary and etoposide, which
is a control compound to induce >98%
DNA damage (Figure 1). Propidium iodide
is included to provide a total cell count.
This simple detection method utilizing
the superior Chromeo 488 ﬂuorescenceand propidium iodide stain makes the kit
ideal for both high-throughput and highcontent screening in all cells (Figure 2).

% Apoptosis

Start screening compounds today!
Screening treatments for the inductionof DNA damage and apoptosis has neverbeen easier with our DNA Damage Assay.
For more information about this assay
and our other DNA Damage and Repair
assays, please give us a call.

Speciﬁcity for low background
The assay relies on a high-quality rabbit
polyclonal phospho-Histone H2AX

1,000,000

1,000,000
900,000

900,000

800,000

800,000

700,000
600,000
500,000
400,000
300,000

700,000
600,000
500,000
400,000
300,000

200,000

200,000

100,000

100,000

080

Figure 3: Merged image of etoposide-treated HeLa cells.
HeLa cells stained only with propidium iodide (PI) appear
red, while those stained with both PI and phospho-H2AX
detected with Chromeo 488 appear more yellow.

Propidium Iodide Fluorescence Intensity

How does the assay work?
The DNA Damage Assay is a cell-based
assay conducted in 96-well plates. Each
kit includes all the reagents necessary fortwo 96-well plates. Cells are grown andtreated in the plate, then ﬁxed and incubated with the phospho-Histone H2AX
(Ser 139) apoptosis marker antibody.
Next, a short incubation with Chromeo™
488 secondary antibody is performedfor detection. The cells are then washedand stained with propidium iodide andread on a ﬂuorescent plate reader.

Propidium Iodide Fluorescence Intensity

When cells are exposed to ionizing radiation or certain treatment compounds,
double-stranded DNA breaks are createdthat rapidly result in the phosphorylationof the histone variant H2AX at serine 139.
Because 0.03% of chromatin becomes
phosphorylated at the serine 139 residueof H2AX per every double-stranded
DNA break, phospho-H2AX is a sensitivemarker for DNA damage, making it auseful marker for apoptosis1. By assaying for H2AX phosphorylation, Active
Motif’s new DNA Damage Assay makes
screening compounds for their ability
to induce DNA damage or apoptosis
sensitive, accurate and very simple!

(Ser 139) antibody which is subsequently
detected with the Chromeo 488 Goat
anti-Rabbit secondary. Our Chromeo ﬂuorescent secondaries are highly speciﬁcwith extremely low background, so yourresults are always accurate (Figure 3).

0

100,000

200,000

300,000

400,000

500,000

600,000

Chromeo™ 488 Fluorescence Intensity

700,000

800,000

900,000

1,000,00000100,000

200,000

300,000

400,000 500,000 600,000 700,000
Chromeo™ 488 Fluorescence Intensity

800,000

900,000

1,000,000

Figure 2: IsoCyte™ single cell analysis of phospho-H2AX detected with Chromeo 488 secondary in HeLa cells.
Untreated HeLa cells (left panel) are detected mostly in the propidium iodide channel while etoposide-treated cells areseen in the green (488 nm) channel indicating a high degree of DNA damage.

60
40
20
0

Control

TreatedFigure 1: Percent induction of apoptosis in HeLa cells.
Levels of phospho-H2AX measured in untreated (control)and cells treated with etoposide. Averages of quadruplicates are shown.
REFERENCES

1. Rogaku, E.P. et al. (1998) J. Biol. Chem. 273(10):5858-5868.

2

Product

FormatCatalog No.

DNA Damage Assay (Fluorescent)2 x 96 rxns

18030

Toll Free — 1 877 222 9543

October 2007 • volume 8 • number 4

SUMOylation & DNA Methylation

Simpliﬁed Study of SUMOylation Effects with SUMOlink™
SUMOlink™ Kits are a simple, effective method for generating SUMOylated
proteins in vitro. These fast, efﬁcient and robust assays contain all necessary
reagents for SUMOylation of target proteins, and include positive andnegative controls to ensure your success.
The SUMOlink method
SUMOlink™ Kits enable you to easily
perform and detect post-translational
modiﬁcations by SUMO (small ubiquitin-like modiﬁer). With SUMOlink, you
simply add the assay components to a
microcentrifuge tube with your protein
of interest. After a 3-hour incubation,
the reaction is stopped and results canbe analyzed by Western blot (Figure 1).
With the kit’s p53 antibody and either
SUMO-1 or SUMO-2/3 antibodies, you
can easily see the extent to which yourtarget protein has been SUMOylated.

Everything you need to study SUMO
The kits contain E1 activating and E2
conjugating enzymes along with wildtype and mutant SUMO-1 (SUMO-1 Kit)
or SUMO-2 and -3 proteins (SUMO-2/3
Kit). Antibodies for SUMO-1 or SUMO2/3 modiﬁcations, as well as control p53
protein and antibody, are included. For
complete information, please give us acall or visit us at www.activemotif.com.

MW

2

3

1

2

3

98
64
50
36
30
16
p53 antibody

SUMO-1 antibody

Figure 1: Speciﬁc SUMO-1 labeling of p53 by SUMOlink.
Western blot analysis of in vitro SUMOylation of p53
protein by wild-type and mutated isoforms of SUMO-1:
The two Western blots were incubated with p53 antibody(1:5000 dilution) and SUMO-1 antibody (1 :4000 dilution)
Lane 1: Wild-type SUMO protein conjugation reaction.
Lane 2: Mutated SUMO protein conjugation reaction.
Lane 3: No p53 control protein used in conjugation.

Product

FormatCatalog No.

SUMOlink™ SUMO-1 Kit

20 rxns

40120

SUMOlink™ SUMO-2/3 Kit

20 rxns

40220

Easily Compare CpG Methylation of Your Samples
MethylCollector™ provides a fast, efﬁcient protocol for isolating andcomparing CpG-methylated DNA in cell or tissue samples. The kit uses a
recombinant Methyl-binding protein (MBD2b) to capture DNA, rather thanantibody-based immunoprecipitations, which improves assay sensitivity.
MethylCollector advantages• Fast and easy protocol completed
in less than 4 hours• Flexible – enables detection
from 5 ng to 1 µg of DNA• Positive control DNA and PCRprimers ensure success

DNA fragments with little or no
methylation. Ready-to-use methylated
DNA is then eluted. MethylCollector ishighly efﬁcient, enabling analysis of themethylation state of any speciﬁc locus
on genomic DNA isolated from less than800 cells (~5 ng DNA, Figure 1).

The MethylCollector method
In MethylCollector, His-tagged recombinant MBD2b protein speciﬁcally binds
to CpG-methylated DNA fragments
prepared by enzymatic digestion orsonication. These protein-DNA complexes are captured with nickel-coatedmagnetic beads and washed with astringent high-salt buffer to remove

MethylCollector applications
Speciﬁc isolation of methylated DNA by
MethylCollector enables many powerfulapplications, including rapid screening of
the methylation status of multiple lociin tumor tissue or cells and detectingchanges in DNA methylation in otherwww.activemotif.com

1

Positive control
DNA (A431)
M

1

2

3

4

5

Negative control
DNA (HeLa)
6

M

1

2

3

4

5 6

Figure 1: CpG-methylation in HeLa and A431 cells.
Increasing amounts of genomic DNA from A431 (positive
control DNA) and HeLa (negative control DNA) cells
were assayed using MethylCollector and analyzed by
PCR using the kit’s control primers, which amplify a locus
that is not methylated in HeLa but highly methylated
in A431. MethylCollector-enriched A431 DNA generates
robust signals proportional to the amount of starting
material (left panel, lanes 4 to 6). No signal is observedin HeLa DNA (right panel, lanes 4 to 6) or in samples
where His-MBD2b protein was omitted from the binding reaction (left and right panels, lanes 1 to 3). Taken
together, these results indicate that MethylCollector
specifically enriches for methylated DNA fragments, andthat this enrichment is due to the presence of the kit’s
His-MBD2b protein.

situations, such as normal cellular differentiation and aging. To ﬁnd out more,
please visit www.activemotif.com.

Product

FormatCatalog No.

MethylCollector™

25 rxns

55002

3

Histone Purification

NEW: Isolate Pure, Separate Fractions of Histone H2A/H2B and H3/H4
While Preserving Post-translational Modiﬁcations
Now you can easily purify histones and further separate the fractions of core histones from any cell culture or tissue
sample while maintaining post-translational modiﬁcations like acetylation, methylation and phosphorylation states.
Histone puriﬁcation made easy
Active Motif’s Histone Puriﬁcation Kit
enables you to isolate core histones
from any cell culture or tissue sample(Figure 1). The core histones may be puriﬁed as one total population containing
H2A, H2B, H3 and H4, or further puriﬁed
into separate fractions of H2A/H2B andH3/H4 (Figure 2).
MW

Log.

Mitotic
ArrestedMW

SodiumButyrateCore Histones

98
64

98

50

64
50

36

36

30

H3
H2B
H2A
H4

16

30
H3
H2A/H2B

16

H4

6

6

4

Figure 1: Core histones isolated from cells and tissue.
Total core histones were isolated from HeLa cells (leftgel) and rat brain tissue (right gel).
MW
98
64

investigating the transcriptional activity of a gene ofinterest. Our proprietary
buffer system is optimized
to maintain these modiﬁcations while separating
the histones into pure
fractions (Figure 3).

Acid Extraction Method
SodiumButyrate

Mitotic
Arrested

Log.

Active Motif Purification Method
SodiumButyrate

Mitotic
Arrested

Log.
Acetylation

Acid Extraction Method
Log.

Mitotic
Arrested

SodiumButyrate

Active Motif Purification Method
Log.

Mitotic
Arrested

SodiumButyrate

Methylation

Active Motif Purification Method
Acid Extraction Method
How does it work?
µg
8.0
4.0
2.0
1.0
0.5
8.0
4.0
2.0
1.0
0.5
Unlike histone puriﬁcationby acid precipitation, ourPhosphorylation
method utilizes puriﬁcation resin and a series of
Figure 3: Post-translational modiﬁcations preserved.
proprietary elution buffers Acetylation, methylation and phosphorylation states are preserved as well or
better with the Histone Purification Kit vs. a straight acid precipitation method.
to isolate very pure histone
used with the Chromatin Assembly Kit
fractions. The resin has a high binding(page 5) to enable the generation of
capacity for histones, so core histones
chromatin that very closely resembles
may be isolated from small cell culture
native chromatin for functional assays
samples up to grams of tissue. Sequen(Figure 4).
tial elution steps let you collect the core
histones in either one step containing
all core histones or into two separate
populations: one enriched for H2A andH2B and a second fraction containing>90% pure H3 and H4.

50
36
30

16

H2B
H2A

H3
H4

Figure 4: Chromatin assembled with puriﬁed histones.
Histones were purified from HeLa cells and used in the
Chromatin Assembly Kit. The ordered spacing of nucleosomes was confirmed and analyzed by agarose gel.

Better substrate for downstream assays
Core histones isolated by the Histone
Puriﬁcation Kit method are highly pureand suitable for substrates for downstream assays. Puriﬁed histones can be

Try it today!
Call or visit us at www.activemotif.com
to ﬁnd out more about the Histone
Puriﬁcation Kit and related products thatwill beneﬁt your research.

1

2

6
4

Figure 2: Separate H2A/H2B and H3/H4 fractions.
H2A/H2B and H3/H4 fractions isolated from HeLa cells.

Preserve important modiﬁcations
Preserving post-translational modiﬁcations like acetylation, methylation andphosphorylation states is critical when

4

What’s in the kit?
Each kit contains all the necessary equilibration, neutralization and elution buffers plus a reusable puriﬁcation column
for 10 histone puriﬁcations.

Product

FormatCatalog No.

Histone Puriﬁcation Kit

10 rxns

40025

Toll Free — 1 877 222 9543

October 2007 • volume 8 • number 4

Chromatin Assembly

Create Chromatin In Vitro to Reveal Regulatory Mechanisms
Active Motif’s Chromatin Assembly Kit enables you to generate chromatin
in vitro from your linear or supercoiled DNA. It yields chromatin that closely
mimics natural in vivo chromatin, so you can discover which histone modiﬁcations and associated proteins are crucial to regulation of your target.
A simple way to generate chromatin
Now you can investigate regulation ofyour gene of interest in its native form
by assembling it into chromatin using
Active Motif’s Chromatin Assembly Kit.
The kit includes all the recombinant
proteins, core histones, buffers andATP-utilizing factors needed to generate
chromatin in vitro from your DNA, andalso to verify successful assembly. Highquality chromatin with more than six
regularly spaced nucleosomes is made byadding the supplied components to 1 µgof your linear or supercoiled DNA, thenincubating for 4 hours. A simple partial
enzymatic digestion of the resulting
chromatin conﬁrms the ordered spacingof nucleosomes (Figure 1).
M

1

2

3

assembled state often cannot reveal themechanism of transcriptional activation
or repression with the associated factors
and relevant histone modiﬁcations.
However, properly assembled chromatinwith regularly ordered nucleosomes is an
excellent substrate for subsequent assays such as in vitro transcription assays,
histone acetyltransferase (HAT) assays(Figure 2) and ChIP (Figure 3).

- +

p53

p300
p53

4

H3
H2B
H2A
H4

1

2

HAT Assay
Figure 2: HAT assay using assembled chromatin.
Chromatin was in vitro assembled with p300 and acetylCoA, in the presence (+) or absence (-) of p53 at 30°C for
30 minutes. The sample was then analyzed on an
18% SDS-PAGE gel and visualized following fluorography.
Figure 1: Enzymatic digestion of assembled chromatin.
Chromatin assembled from 1 µg samples of circular DNA(Lanes 1 & 2) and linear DNA (Lanes 3 & 4) were digested
for 2 and 4 minutes, respectively, deproteinated, phenol/
chloroform extracted and run on an agarose gel. Eachsample type resulted in regularly spaced nucleosomes.

Why bother to make chromatin?
When DNA sequences are assembled
into chromatin by ATP-dependent
factors, the resulting structure closely
resembles the natural chromatin conﬁguration. DNA that is in either a bare or un-

www.activemotif.com

More natural for true results
Now you can use histones puriﬁed from
actual samples with the Histone Puriﬁcation Kit (page 4) to create chromatin thatclosely mimics the in vivo form, so you
can conﬁrm results and be certain about
which factors are critically involved inregulation of your target sequence.

Chromatin Assembly Kit advantages• Generate chromatin from linear orsupercoiled DNA• ATP-dependent method results
in an extended array of regularly
spaced nucleosomes• Easy protocol – simply incubate thekit components with your DNA• Produces an excellent substrate forvarious gene regulation experiments

Input
AP-1

anti-hexahistidine- + - +

Figure 3: ChIP of in vitro assembled chromatin.
In vitro ChIP was performed using in vitro-assembled
chromatin containing an AP-1 binding site in thepresence of p300 and acetyl-CoA, with or withouthexa-histidine-tagged AP-1. The ChIP was performed
with a hexahistidine antibody and PCR was conductedwith primers flanking the AP-1 binding site.

Complete kit ensures your success
The kit includes recombinant h-NAP-1
chaperone protein, ACF assembly complex, HeLa core histones, and the buffers
and enzymes necessary to generateassembled chromatin from your input
DNA. Control Supercoiled DNA is also
provided. To verify that your chromatin
assembly reaction has been successful,reagents are provided to perform aneasy partial enzymatic digestion to visualize the regular spacing of nucleosomesby agarose gel electrophoresis (Figure 1).
Start assembling chromatin now!
The Chromatin Assembly Kit makes it
easy for you to assemble chromatin on
your sequence of interest to investigate
gene regulation in a context that closely
resembles in vivo chromatin. For more
information, please give us a call.

Product

FormatCatalog No.

Chromatin Assembly Kit

10 rxns

53500

5

Chromatin Immunoprecipitation

Improved ChIP-IT™ Express Greatly Reduces Background
Active Motif has improved its ChIP-IT™ Express Kits by greatly reducing
the background, which improves your results and enables you to use lessstarting material than ever before. In addition, the provided magnetic beads
have made it possible to streamline the protocol so you can get results inhalf the normal time with much less sample manipulation. And, ChIP-IT
Express makes it easy to perform ChIP on many samples at the same time.
ChIP-IT Express advantages• No more need for pre-clearing,
blocking or DNA puriﬁcation steps• Reduced background• High throughput compatible• Dramatically reduced hands-on time

table result. Conventional ChIP requires
at least 2 million cells as starting material, which can be problematic with somecell lines. At the least, growing this manycells is labor intensive. Active Motif’s
improved ChIP-IT Express Kits, however,
have been optimized to provide superior
target gene enrichment, resulting in
unmatched sensitivity. Using ChIP-IT
Express, it is routine to perform ChIP onmaterial from as few as 750,000 cells andthe kits have even been shown to workwith as few as 12,500 cells (Figure 1)!

The most efﬁcient ChIP enrichment kit
ChIP is an enrichment technique, nota puriﬁcation method. Thus, the less
efﬁcient your enrichment, the higher the
sample background and the more material you will need to obtain an interpre-

25,000 cells

Neg IgG

RNA pol II

Neg IgG

RNA pol II

Neg IgG

RNA pol II

Neg IgG

RNA pol II

GAPDH Primers

12,500 cells

Figure 1: ChIP-IT Express works with 12.500 cells.
ChIP-IT Express was performed in duplicate on decreasing amounts of sonicated HeLa cell chromatin. Twoµg of RNA pol II and Neg IgG antibody was used for IP. GAPDH PCR primers were used to analyze theimmunoprecipitated DNA. Using the improved ChIP-IT Express reagents and protocol, positive ChIP data
was obtained from as few as 12,500 cells.

Product

FormatCatalog No.

ChIP-IT™ Express

25 rxns

53008

ChIP-IT™ Express Enzymatic

25 rxns

53009

ChIP-IT™ Protein G Magnetic Beads

25 rxns

53014

ChIP-IT™ Control Kit – Human

5 rxns

53010

ChIP-IT™ Control Kit – Mouse

5 rxns

53011

ChIP-IT™ Control Kit – Rat

5 rxns53012

Ready-to-ChIP HeLa Chromatin

10 rxns

53015

6

The magnetic bead advantage
The ChIP-IT Express magnetic beads haveless background than standard agarose
beads, and this means pre-clearing andblocking steps are no longer necessary.
The magnetic pull-down occurs in justseconds, and re-formulated buffers
allow steps to be combined and DNA
puriﬁcation to be eliminated. ChIP-IT
Express is available in both sonicationand enzymatic shearing formats.
Positive controls ensure success
Because interpreting ChIP data can be
difﬁcult, Active Motif has developed acomplete set of controls to help you
understand your results and troubleshoot your assays. To provide you with
controls that are appropriate for yourresearch, we removed the human-only
controls from ChIP-IT Express Kits andnow offer human, mouse and rat ChIP-IT
Control Kits separately. These provide
positive and negative control antibodies
and species-speciﬁc primers, PCR buffer
and a convenient 10X DNA loading dye
so your PCR reactions are gel-ready. All
reagents are quality control tested andvalidated to ensure your ChIP assay is
working properly. In addition, we alsooffer convenient Ready-to-ChIP HeLa
Chromatin, so you can be certain thatthe only variable in validating a new
antibody for ChIP is the antibody itself.
Try the best ChIP kit today
For additional information on the new
and improved ChIP-IT Express Kits, visit
our website at www.activemotif.com.

Toll Free — 1 877 222 9543

October 2007 • volume 8 • number 4

Co-Immunoprecipitation

Simpliﬁed Co-Immunoprecipitation of Intact Nuclear Complexes
The Nuclear Complex Co-IP Kit simpliﬁes co-immunoprecipitation of intact
nuclear protein complexes because its reagents have been optimized for
preparing nuclear extracts and immunoprecipitating DNA-bound proteins.
Co-Immunoprecipitation (Co-IP) is often
used to study protein/protein interactions. In Co-IP, a ﬁrst antibody is used
to immunoprecipitate a target antigen,which also co-precipitates any bound,interacting proteins. These are then detected by Western blot using antibodies
targeted against the interacting proteins.
However, traditional Co-IP methods are
not optimal for studying DNA-binding
proteins because their complexes arevery fragile, so are frequently disrupted
during extraction. In addition, thesecomplexes can be altered by the salt
and detergent composition of theimmunoprecipitation buffers, which canfurther complicate their analysis.

Perform Co-IP of DNA-bound proteins
To overcome these problems, the Nuclear Complex Co-IP Kit contains extraction
reagents that were designed to help
maintain nuclear protein complexes. The
kit’s extraction process provides a simple,
effective method for isolating intact
protein complexes contained in nuclear
compartments of the cell, speciﬁcallythose previously bound to DNA. Afterisolation of the intact complexes, thesupplied Co-IP reagents were designed
so that you can vary the stringency of
the Co-IP buffers. This improves yourresults by making it easy for you to
study any protein complex, whether its
members are tightly or weakly bound.

“The Nuclear Complex Co-IP Kit improves co-immunoprecipitation of
DNA-binding proteins by providing extraction and immunoprecipitation
components that are optimized to maintain nuclear protein complexes.”
Cell nucleus containing the twointeracting proteins of interest

1

2

3

Nuclear extraction

Add antibody
binding beads

Add antibody directed against
one of the proteins of interest

4
Lane 1. Mol. Weight Marker
Lane 2. Extract
Lane 3. negative IP (no ab)
Lane 4. Co-IP

Western blot analysis of the immunoprecipitated proteinsusing an antibody directed against the second protein of interest

Wash & collect theimmunoprecipitated proteins

Immunoprecipitate theproteins of interest

Figure 1: Flow chart of the Co-Immunoprecipitation process.

Product

FormatCatalog No.

Nuclear Complex Co-IP Kit

50 rxns54001

www.activemotif.com

250
1

2

3

Figure 2: Analysis of p33 in the RNA pol II complex.
HeLa cells were grown to confluence on 100 mm plates
and nuclear extracts were prepared using the kit’s
extraction reagents. For IP experiments, the stringencyof the IP High Buffer was increased by supplementing
with NaCl and Detergent. 100 µg of nuclear extract wasused per IP reaction and incubated with either 2 µg
p33 antibody or no antibody. Following the IP, Western
blot analysis was performed using RNA pol II mouse
mAb at 0.1 µg/ml followed by anti-mouse HRP at 1:1000.
Detection of the p33/RNA pol II complex by the RNA
pol II antibody (lane 3) demonstrates that the Co-IP
was successful in maintaining the protein complex. The
input HeLa extract (lane 1) was run as a control for theWestern blot using 0.1 µg/ml RNA pol II.
Lane 1 Western blot control
Lane 2 Negative Control (no antibody used in IP)
Lane 3 Co-IP: IP using p33/WB using RNA pol II

Easy to optimize for ideal stringency
With the Nuclear Complex Co-IP Kit,
nuclear extracts are prepared using
low-salt buffers and enzymatic shearing.
The low-salt buffers keep the protein
complexes intact, while digestion
gently releases them from the DNA.
Immunoprecipitation is then carriedout to detect the bound proteins. The
kit contains high- and low-stringency IP
buffers, as well as salt and detergent.
Addition of salt and detergent is ideal
for robust protein/protein interactions
because higher stringency reduces background. However, as unstable protein
complexes may not withstand highstringencies, the kit makes it simple tooptimize the stringency as required foreach particular protein complex.
Find the complex members you missed
The Nuclear Complex Co-IP Kit offers
you a simpler, more ﬂexible alternative
than traditional co-immunoprecipitation
methods. To ﬁnd out more about how it
can help you, please give us a call or visit
us on the web at www.activemotif.com.

7

Automated Text Mining

Learn More Faster With AlmaKnowledgeServer 2
AKS2 lets you dig deeper than othersearch tools. It utilizes advanced text
mining to automatically examine thethousands of new abstracts added to
PubMed every day. AKS applies rulesbased analysis to determine which words
are bioentities, such as genes, chemicals, drugs, diseases, etc. This content
is added to the AKS database, which issearched to reveal relationships between
bioentities and their relevance. A varietyof graphical tools make it easy to quickly
ﬁnd the information that is important toyou (Figures 1-4). AKS can even discoverlinks that have not been published.
The new AKS v2.1 includes:• Automatic abstract alerts via email• PubChem compound structures• Publication trend analyses• Improved export tools and graphics

Figure 2: Summary results for the gene/protein CDK2, which shows the relevance of co-occurring bioentities.

01/2006

16370931| Expert Opin Investig Drugs 2006,Jan,01;15(1):23-38

Wesierska-Gadek Józefa ; Schmid Gerald

The inhibition of cyclin-dependent kinases (CDKs) represents a novel approach to the therapy of human malignancies. Already in clinical trials,
cells and inhibit their cell-cycle progression. Interestingly,

recently developed CDK inhibitors very efficiently target the rapidly proliferatingsome CDK inhibitors additionally affect the stability and activity of the tumour-suppressor protein p53, thereby enhancing their antiproliferative action
towards

cells. Considering the fact that the p53 protein is mutated or inactivated in approximately 50% of all human

CDK inhibitor therapy could differ between

inhibition questioned the central role ofcells can proliferate despite, the efficacy ofcells depending on their p53 status. Moreover, recent reports demonstrating that somein the cell-cycle control and suitability of

as a therapeutictarget; however, the p53 activation that is mediated by CDK inhibitors could be essential for the efficacy of CDK inhibitors in therapy of CDK2-

Why not give it a try? For a free 7-day
demo, visit www.activemotif.com/aks.

. Furthermore, there is also reason to believe thatindependent

inhibitors could be used for another purpose, to protect normal cells

from the effects of chemotherapy.

Figure 3: Abstracts are displayed with your choice of bioentity types highlighted, so you can quickly ﬁnd relevant
information.
08/2003
12888290| Trends Cell Biol 2003,Aug,01;13(8):396-9

Grim Jonathan E ; Clurman Bruce E(
that control
some

, and

) regulates diverse aspects of the mammalian cell cycle. Most
activity has received much attention as a target forcells can proliferate without

activity questions the essential role ofcells contain mutations in the pathways

therapy. However, a recent report demonstrating thatin cell-cycle control, as well as its suitability as atherapeutic target.

Hide/Show this panel

Figure 1: Chemical structure and trend analyses.

Figure 4: AKS includes a powerful visual environment to map relationships between bioentities.

Online Access to AKS v2.1 is Now Available at a Low Introductory Price!
With the introduction of AKS v2.1, Active Motifis pleased to announce that AKS is now available
as an online subscription service. This is an ideal,
low-cost solution that untethers AKS from yourcomputer, making it available everywhere.

Through the end of 2007, an online subscription
to AKS is available for just $100/€99 per month,a substantial discount from the normal price of$150/€150. To place your order, please contact
your local Active Motif ofﬁce.