Chromatin Immunoprecipitation

ChIP-IT® Express
RNA ChIP
Sequential ChIP
High-throughput ChIP
ChIP-validated Antibodies
ChIP Controls & qPCR Primers

the innovation leader in ChIP products
Active Motif’s extensive line of ChIP-IT® Express Kits andreagents utilize the many innovations we have developed
to make ChIP faster, simpler and more reproducible.
We also offer large, rapidly growing collections of
ChIP-validated antibodies, qPCR primer sets andcontrols. Additionally, our Epigenetics Services
can perform the latest ChIP techniques for you.

EpiShear™ ChIP Sonicators
ChIP DNA Purification
ChIP-Seq Services
Enabling Epigenetics Research

The Innovation Leader in ChIP products
Active Motif has consistently been the leader in introducing innovations that make ChIP faster, simpler and more
reproducible. We were the first to introduce magnetic beads into a ChIP kit, the first to offer enzymatic chromatin
shearing and, building on these improvements, the first with kits for performing sequential, high-throughput andRNA ChIP. We continue to expand our ChIP product line by focusing on providing a complete selection of provenreagents and equipment for ChIP, including the largest collection of antibodies validated for ChIP, ChIP-Seq and
ChIP-chip. In addition, we recognize that as ChIP has transitioned to a genome-wide technology, not every lab has
the equipment or know-how required to perform the latest techniques. To enable our customers to keep up in
this rapidly evolving field, we offer end-to-end Epigenetic Services such as ChIP-Seq, MeDIP-Seq and ChIP antibody
validation. Customers can send samples to Active Motif and weeks later receive fully analyzed data.
ChIP-IT® Express – chromatin immunoprecipitation made easy
Chromatin immunoprecipitation (ChIP) isa powerful tool for studying protein/DNA
interactions. It is the most-widely citedepigenetic technique and can be used togenerate both single-gene and genome-wide
data. However, ChIP can be technically challenging and yield results that are difficult tointerpret. In order to minimize the challenges
associated with ChIP and help ensure yoursuccess, Active Motif offers its ChIP-IT®
Express Kits. Each kit features a streamlined
protocol that reduces the amount of sample
required and minimizes the background signal obtained. Consequently, ChIP-IT Express
Kits are the most widely published ChIP kitsavailable (Figure 1).
In the ChIP-IT Express method, intact cells
are fixed using formaldehyde, which crosslinks and therefore preserves protein/DNA
interactions. The DNA is sheared into small
fragments using either sonication or enzymatic digestion. The protein/DNA complexes
are then immunoprecipitated using antibodies directed against DNA-binding proteins or
histone modifications. Following immunoprecipitation, the cross-linking is reversed andthe enriched DNA fragments are analyzed by
qPCR or in whole-genome applications suchas ChIP-Seq (Diagram 1).

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Figure 1: ChIP-IT Express Kits are cited in more publications than any other manufacturer’s ChIP kit.
Trade name searches were performed on HighWire Press (http://highwire.stanford.edu/) for ChIP kit journal citations.
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gifk\`e&;E8Zfdgc\o\jXe[nXj_lj`e^DX^e\k`Z9\X[j%Diagram 1: Schematic of chromatin immunoprecipitation using ChIP-IT Express.
In ChIP, protein/DNA interactions are fixed, and the DNA is sheared and precipitated using an antibody. After crosslink reversal, the DNA is analyzed, usually by qPCR, to determine which genes were bound by the protein of interest.

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ChIP-IT Express is a faster, more streamlined method for successful ChIP
ChIP-IT Express Kits improve on traditional
ChIP by reducing or eliminating several timeconsuming steps. Our kits were the first to
utilize magnetic beads, which have 4much
lower background binding than traditional
agarose beads. This reduced background has
made it possible to eliminate some of thesteps required in more conventional protocols like pre-clearing and blocking. Washingis much easier because the spin steps havebeen replaced by rapid magnetic pull-down.
Collectively, all of the improvements included in ChIP-IT Express give you the capability
to perform your reactions in PCR tubes witha multi-channel pipettor, which reduces youreffort and improves consistency (Figure 2).

Figure 2: Multiple sample ChIP using ChIP-IT Express.
Washing the magnetic beads is fast and easy because the pellet forms against the side of the tube
above the level of the supernatant. This speeds the procedure, eliminates sample loss and enables
you to ChIP multiple samples in 8-well PCR tubes using a multi-channel pipettor.

(A bar magnet is included with all ChIP-IT
Express Kits, but you can also use commercially available magnetic stands.) For more

Neg IgG

RNA pol II

ChIP-IT Express advantages

Neg IgG

ChIP is an enrichment technique, not a
purification method. Most marks of interest
exist throughout the genome, but in different relative proportions. Consequently,
meaningful results require efficient enrichment. Conventional ChIP requires at least
two million cells as starting material, which
can be problematic with some cell lines, andlabor-intensive even in the best case. ChIP-IT
Express Kits have been optimized to provide
superior target gene enrichment, resulting in
unmatched sensitivity. The kits can routinely
produce excellent results with chromatinfrom as few as 100,000 cells (Figure 3).

RNA pol II

Efficient ChIP enables you to use fewer cells

complete information on ChIP-IT Express
Kits, please give us a call or visit our
website at www.activemotif.com/chip.

Figure 3: ChIP-IT Express works with 100,000 cells.
ChIP-IT Express was performed in duplicate onsonicated HeLa cell chromatin (1.0 x 105 cell equivalents
per ChIP). Two µg of RNA pol II and Neg IgG antibody
was used for IP. GAPDH PCR primers were used to analyze the immunoprecipitated DNA. DNA isolated with
RNA pol II antibody generated more product than thenegative control IgG. These results demonstrate that
positive ChIP data was obtained from 100,000 cells.

• Reliability – ChIP-IT Express Kits are themost widely cited ChIP kits
• Faster procedure – magnetic capture ismuch faster than centrifugation• Simplified wash steps – improved
consistency and dramatically reduced
hands-on-time• No pre-blocking needed – magnetic
beads are inert and exhibit low bindingof proteins• Less effort required – our kits arecompatible with multi-channel
pipettors to streamline wash steps

Shear chromatin with your choice of sonication or enzymatic digestion
Chromatin shearing is most commonly doneby sonication. However, sonication can bedifficult to optimize due to emulsificationand overheating of the sample. Because ofthis, or if you don’t have a sonicator, Active
Motif has developed a robust, user-friendly
method to shear chromatin by enzymatic
digestion (Figure 4). Because Active Motif
offers both ChIP-IT Express and ChIP-IT
Express Enzymatic Kits, you can choose
whichever shearing method you prefer.
(Stand-alone shearing kits are also available.)

Figure 4: Analysis of enzymatically sheared DNA.
HeLa cells were fixed for 10 minutes with formaldehyde and then chromatin was prepared using the
Enzymatic Shearing Kit protocol & reagents. Chromatin
was sheared with the Enzymatic Shearing Cocktail for
5, 10 & 15 minutes before the reaction was stopped.
Lane 1: 100 to 1000 bp ladder.
Lane 2: Unsheared HeLa DNA.
Lane 3: HeLa DNA treated for 5 minutes.
Lane 4: HeLa DNA treated for 10 minutes.
Lane 5: HeLa DNA treated for 15 minutes.

1000 bp
500 bp100 bp
1

2

3

4

5

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The Innovation Leader in ChIP products
ChIP-validated antibodies – guaranteed to work in ChIP
Active Motif is committed to providing
the highest quality antibodies for studying
chromatin and the biology of the nucleus,
especially antibodies validated for use inchromatin IP (Figure 5). We specialize in
manufacturing antibodies against histones,
histone modifications and chromatin proteins. Because ChIP is an extremely challenging technique, only antibodies of the highest

quality that recognize the target protein in
its native, chromatin-associated context will
work. Unlike other antibody “suppliers”, wemanufacture and test our own antibodies, so
we guarantee they will perform as specified.
Simply let us know if you are not satisfied.
For a complete listing of our ChIP-validated
antibodies, please visit our website atwww.activemotif.com/chipabs.

Figure 5: ChIP of methylated histone H3.
ChIP was performed using chromatin prepared frommouse embryonic stem cells (left lane) or mouseerythroleukemia cells (right lane) with H3K4me2 pAb(Cat. No. 39141). PCR primers specific for b-globin wereused to amplify a 210 bp region of the promoter.

ChIP-IT Control Kits help ensure successful ChIP and simplify antibody validation
The ChIP-IT Control qPCR Kits include a
positive control antibody, a bridging antibody to enhance binding affinity of mousemonoclonal antibodies, a negative control
antibody to evaluate non-specific binding,and species- specific positive and negative
control qPCR primers. The positive and negative control primers allow you to show that
the ChIP worked and to establish backgroundbinding levels within the same ChIP. Negative
primers can be a more relevant control than
IgG as different antibodies will have different
background binding capacities. (Figure 6).
Our original ChIP-IT Control Kits are designed
for use in endpoint PCR. Each kit provides
positive and negative control antibodies, apositive control PCR primer set as well as(%-=7F:>#(
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ChIP is a method of enrichment of DNA
bound by a particular protein, not a complete purification of the protein-bound DNA.
As a result, ChIP reactions are unavoidably
contaminated with non-specifically captured
chromatin. This can lead to false positive
PCR products that make data interpretationdifficult. To solve this problem, Active Motif
offers species-specific (human, mouse andrat) ChIP-IT Control Kits for real-time or endpoint PCR. Incorporation of all of the controls into your ChIP experiment allows you
to evaluate your success at multiple steps.
The control kits help confirm the chromatin
preparation and immunoprecipitation procedures worked properly and enable you toassess the quality of your test antibody and
PCR reactions.

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Figure 6: qPCR analysis of human myeloma LP1 ChIP.
ChIP was performed on 5 µg of chromatin fromhuman LP1 cells using the ChIP-IT Express Kit andthe positive & negative control antibodies and thepositive & negative control qPCR primers that aresupplied in the ChIP-IT Control qPCR Kit – Human.

a PCR buffer/DNA loading dye that makes
your PCR reactions ready to run on the gelstraight out of the thermocycler. Please visit
www.activemotif.com/chipcontrols for details.

Convenient collections of species-specific qPCR primers and Ready-to-ChIP Chromatinfying enzymes, RNA Pol II and methylatedDNA. To see the full list of over 35 available
ChIP Control qPCR Primer Sets and their targets, visit www.activemotif.com/chipprimers.
We also offer Ready-to-ChIP Chromatinfrom a number of ENCODE cell lines that are
optimally sheared by sonication and ChIPvalidated. The chromatin can be used with all
ChIP-IT Express Kits and controls, so you canbe certain that the only variable when validating a new ChIP antibody is the antibody
itself. See www.activemotif.com/chipready.

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8_dZ_d];l[dji:[j[Yj[Z%'&&&Y[bbiAs detailed above, ChIP data interpretation ishighly dependent on the use of controls. But,
identifying appropriate controls is a challenging undertaking. Accessing this informationin the published literature is time consuming
and may not be relevant to your cell system.
To help, Active Motif Epigenetic Services
has compiled data from thousands of ChIP
experiments and developed human, mouse,rat, Zebrafish, Drosophila and yeast positive
and negative control qPCR primer sets formany common ChIP targets, including histone modifications (Figure 7), histone modi-;nf[h_c[dj';nf[h_c[dj(

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Figure 7: qPCR with positive & negative primer sets.
ChIP was performed in duplicate on HeLa chromatinwith ChIP-IT Express and H3K4me3 pAb (Cat. No. 39159),
then the Human Negative-1 (Cat. No. 71001), Human
ACTB-2 (Cat. No. 71005) and GAPDH-2 (Cat. No. 71006)
Positive Control Primer Sets were used for qPCR.

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Epigenetic Services – ChIP-Seq, MeDIP-Seq and Antibody Validation Services
Genome-wide ChIP applications require
extensive optimizations at multiple steps
including chromatin preparation, antibody
validation, background reduction andsequencing library construction. The investment in time and money is significant andthe quality of the final data is not known
until the entire project is completed. To takethe guesswork out of generating high-quality
data, Active Motif Epigenetic Services offersa variety of genome-wide data generation
and analysis services such as MeDIP-Seq,
ChIP-Seq (Figure 8), and ChIP antibody validation (Figure 9). Customers send cell pellets
and receive fully analyzed ChIP-Seq data.
Active Motif performs the chromatin preparation, antibody validation, ChIP, library generation, sequencing and data analysis. Quality
control steps incorporated at multiple points
in the process ensure high-quality data uponcompletion of the project. For more information on all of the services available, please
visit www.activemotif.com/services.

Figure 8: ChIP-Seq of a specific histone modification.
ChIP–Seq was performed using chromatin from mouse livers and an antibody against H3K4me3 (Cat. No. 39159).
Sequencing was performed on the Illumina GA II using 36 bp single-end reads. Genome alignment was performed
with ELAND and peak calling was performed with MACS. The image above focuses on a 3 Mb window containing a
Zfp gene cluster on chromosome 13. H3K4me3 peaks are present at the start site of all Zfp genes. Gene annotations
run from right to left, therefore TSSs are on the right side of each gene annotation.

Antibody validation for ChIP-Seq and ChIP-chip
The field of epigenetics has grown rapidly
over the past 10 years. Chromatin immunoprecipitation has evolved from a techniqueused to explore transcription factor binding
and epigenetic changes at single genes using
PCR to a technique capable of profiling theentire genome. Active Motif believes that
antibodies should be validated for the applications in which they will be used. In addition to the existing ChIP PCR and specificity
testing we do on every new antibody we
produce, we have undertaken an effort tovalidate all new antibodies in ChIP-Seq and/or ChIP-chip. We are also re-validating ourexisting histone modification and transcription factor antibodies in these applications.
An updated list of our ChIP-Seq and ChIPchip validated antibodies is available atwww.activemotif.com/chipseqabs.

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Figure 9: Flow chart of the steps in our ChIP-chip antibody validation process.
Active Motif makes ChIP antibody validation simple and fast by offering a ChIP antibody validation service, which
we also employ on our own antibodies. Validation is performed using ChIP-chip on tiling arrays, as shown above, orby ChIP-Seq. Active Motif performs all steps of the experiment including data analysis and the performance of theantibody is assessed based on the identification of ChIP-chip peaks.

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The Innovation Leader in ChIP products
Consistent improvements have lead to the development of new ChIP methods
Active Motif has consistently been the leader
in introducing improvements that makechromatin IP simpler and more reproducible.
We were the first company to develop andintroduce a ChIP kit employing magnetic
beads. We were also the first to produce
complete ChIP kits containing all criticalcomponents, including antibody and PCRprimer controls. This lead to the release of
more convenient species-specific control
kits. We were also the first to introduce
enzymatic chromatin shearing. All of these
innovations have made ChIP faster, easierand more successful.

We have strengthened our position as theleader in developing innovative ChIP products by applying these improvements to thecreation of kits for performing high-throughput ChIP, sequential ChIP and RNA ChIP. The
methods provided in these kits will help you
to expand the utility of the ChIP technique.

High-throughput ChIP a reality with ChIP-IT Express HT
If you need to perform many ChIP experiments at once, ChIP-IT Express HT is yoursolution. It combines the time-saving, magnetic bead-based protocol of ChIP-IT Expresswith a high-throughput, 96-well microplatebased format (Figure 10). With ChIP-IT Express
HT you can efficiently process up to 96 ChIP
reactions simultaneously (Figure 11). ChIP-IT
Express HT is compatible with our enzymaticand sonication-based shearing kits, as wellas with our control kits, qPCR primers andReady-to-ChIP Chromatin. For information,please visit www.activemotif.com/htchip.

Figure 10: True high-throughput ChIP.
With magnetic beads, optimized buffers and theefficient plate-based protocol of ChIP-IT Express HT,
you can process up to 96 ChIP reactions at a time.

Figure 11: High-throughput ChIP performed on HeLa
chromatin using ChIP-IT Express HT.
PCR carried out using GAPDH-specific primers. Lanes
1-4: ChIP using 2 µg RNA Pol II antibody. Lanes 5-8:
ChIP using normal mouse IgG as a negative control.
Lane 9: no DNA control. Lane 10: input DNA control.

Identify protein co-localization in vivo using sequential chromatin IP
When performing ChIP experiments to decipher the Histone Code, it is often useful toprove that two different proteins or histone
modifications co-occupy the same site in
the genome. Alternatively, you may want todetermine if a protein coincides with a specific histone modification at the same regulatory element. Sequential ChIP (also called
Re-ChIP) is a technique in which successive
chromatin immunoprecipitation reactions are
performed using two different antibodies,enabling you to assay for the simultaneouspresence of two proteins or distinct histone
modifications at the same genomic region ofinterest. Performing sequential chromatin IP
was technically challenging and difficult, until
now. Active Motif’s Re-ChIP-IT Kit has been
optimized for sequential ChIP (Figure 12),enabling you to perform this difficult technique as easily as our standard ChIP-IT Kits,
and with the ability to use ChIP-IT controls.
Please visit www.activemotif.com/rechipit.

Figure 12: Sequential chromatin immunoprecipitation using Re-ChIP-IT.
The lane numbers are the same in each panel to indicate that the DNA is from the same chromatin sample. The left
panel shows the results of PCR performed on an aliquot of DNA removed from the experiment after the first ChIP
step; the right panel represents PCR results on DNA from chromatin samples after both ChIP steps. For example,
chromatin samples subjected to first ChIP using Mouse IgG as a negative control (lanes 1 and 2 in the left panel) werethen subjected to a second ChIP with an RNA Pol II antibody (lanes 1 and 2 in the right panel). Chromatin samples inwhich Mouse IgG was used as either the first antibody (lanes 1 and 2) or second antibody (lanes 5 and 6) show little
amplification of GAPDH DNA in either the left (first ChIP) or right panel (first and second ChIP). Chromatin samples inwhich the first antibody used was anti-RNA Pol II and the second antibody was anti-TFIIB (lanes 3 and 4) show good
amplification of GAPDH DNA after the second ChIP (right panel) indicating co-localization of RNA Pol II and TFIIB at
the same region of the GAPDH promoter.

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Evidence is building that RNA-directed
processes play a critical role in orchestrating chromatin architecture and epigenetic
memory. Nucleic acids purified from chromatin are 2-5% RNA; these RNAs are non-coding
sequences that play important roles in chromatin structure and transcriptional silencing.
However, characterizing these RNAs using
conventional ChIP techniques is difficult due
to the complexity of chromatin and the large
amount of DNA present in chromatin.
To make the characterization of the role of
RNA in genome regulation possible, Active
Motif has leveraged its expertise in ChIP todevelop the first of its kind kit for RNA-ChIP.
The RNA ChIP-IT Kit was designed to study
RNA-protein interactions in a chromatin context, and optimized to recover RNA for real-time RT-PCR analysis. The kit utilizes magnetic beads for the RNA capture, which improve
results while reducing time and effort.
RNA ChIP-IT uses a modified ChIP protocolthat has been optimized for RNA preservation and recovery. RNA-protein interactions
are fixed with formaldehyde, and chromatin
shearing is combined with DNase treatment
to yield RNA/protein complexes that can be
immunoprecipitated with antibodies to specific proteins. Cross-links are subsequently
reversed; RNA is recovered and again treatedwith DNase to ensure the absence of DNA.
The optimized method is quick and has beensuccessfully used to study several non-coding
RNAs in the chromatin context (Figure 13).
For complete information, please visit us atwww.activemotif.com/rnachip.

_dfkjh[Yel[hoOptimized method to study RNA/protein interactions in a chromatin context
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Figure 13: Percent recovery of RNA-ChIP samples.
The RNA ChIP-IT Kit was used on 10 µg samples of
DNase I-treated HeLa chromatin with 10 µl of Suz12
antibody (Cat. No. 39357) and 2 µg of Normal Rabbit
IgG. Real-time RT-PCR was performed using primers
for the lincRNA SFPQ locus, and the % input recoveries were then calculated and plotted above. Dividing
the input recovery of the Suz12 antibody by that of
the rabbit IgG indicates a 141-fold enrichment of the
lincSFPQ region with Suz12 antibody. These results suggest that the SFPQ lincRNA is a structural componentof the Suz12-containing complex, PRC2.

Rapid, simple purification of DNA from your ChIP experiments

While Phenol/chloroform extraction followed by ethanol precipitation is considered
the gold standard in terms of DNA recovery
and purity, the technique uses dangerous
chemicals and is time consuming. Standard
column-based DNA purification kits havebeen tried as an alternative, but are often
not compatible with some components of
ChIP elution buffers.

In contrast, Active Motif’s Chromatin IP
DNA Purification Kit is a column-based DNA
purification method that is compatible with
our ChIP-IT Express Kits and other ChIP
protocols. Clean up is quick, easy, reliable
and reproducible. The quality of the purified
DNA is equivalent to that isolated by phenol/chloroform extraction and ethanol precipitation (Figure 14). In addition, the kit canbe used to purify methylated DNA enrichedusing Active Motif’s MethylCollector™,
UnMethylCollector™, MeDIP and hMeDIP Kits.
For more complete information, please visit
www.activemotif.com/dnapure.

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8_dZ_d];l[dji:[j[Yj[Z%'&&&Y[bbiThe use of ChIP in combination with a variety of analysis techniques such as qPCR,
ChIP-Seq and ChIP-chip can yield a tremendous amount of information regarding the
distribution of transcription factors andhistone modifications. But, data quality inthese downstream applications is directly
linked to the quality of the ChIP DNA.

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equivalent to phenol/chloroform extraction.
ChIP was performed in duplicate using ChIP-IT Express,
RNA pol II phospho Ser2 mAb (Cat. No. 61083) andchromatin from REC1 cells. After the cross-links werereversed, half of each reaction was purified by phenol/
chloroform extraction followed by ethanol precipitation. The other half was purified using the Chromatin
IP DNA Purification Kit. The purified ChIP DNA wasthen used in qPCR with negative primers that amplifya region in a gene desert (Cat. No. 71001) and positive
control primers that amplify a region in the ACTB gene
(Cat. No. 71003).

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www.activemotif.com

EpiShear™ sonication products make it easier to reproducibly shear chromatin for ChIP
Active Motif’s EpiShear™ line of sonication products
were designed to save you time and effort while
ensuring that you obtain more reproducible results
when shearing chromatin or DNA for use in ChIP, DNA
methylation studies and Next-Gen sequencing.
The EpiShear Multi-Sample Sonicator (Figure 15) is ahigh-intensity cup horn sonicator that can process up
to eight vials (20 µl - 1.2 ml sample per vial) simultaneously. It comes with a sound enclosure and, optionally,a thermoelectric chiller. For complete information,please visit us at www.activemotif.com/cuphorn.
While the EpiShear Probe Sonicator is a compact, economical unit (Figure 16), it is still a fully programmableunit with all the features needed to help ensure reproducible results. It is supplied with a 1/8" microtip probethat enables you to shear one sample at a time (500 µl
to 15 ml). Other size probes are available that expand
its range from 200 µl to 50 ml. For more information,please visit www.activemotif.com/probe.

Figure 15: The EpiShear Multi-Sample Sonicator / Chiller.
The EpiShear Multi-Sample Sonicator / Chiller includes a powerful 750-watt generator
with a keypad and digital display that make it easy to program and monitor sonication.
The unit’s cup horn sonicator is housed in a compact sound enclosure to reduce sonicationnoise. The cup horn can process up to 8 samples simultaneously, which rotate continuously
to ensure all samples are processed equally. It is mounted in an acrylic bath that is filledwith water. The bath and enclosure are plumbed with quick-connect hoses that attach to athermoelectric chiller, which keeps the samples at 2-4°C during sonication.

Both sonicators are controlled by microprocessor units
that offer both programmable and manual operation.
Each has a keypad and digital display that make it easy
to program the amplitude and set the total sonication
time and duration of On and Off pulse cycles. Pulse
intensity can be set from 20-100%, enabling you to
optimize exact parameters for all of your cell types.
The EpiShear Cooled Sonication Platform improves
sample-to-sample reproducibility when using any probesonicator by positioning the tip of the probe at thesame depth in the sample for each sonication. The
platform has been machined out of stainless steel andaluminum, and includes a hand crank, a height counterand a vertical alignment tool. After determining youroptimal settings, you can recreate them every time.
The platform can be used in a sound enclosure oron its base, and includes a Tube Cooler (available formicrofuge, 15 ml and 50 ml tubes). These keep the sample cold during sonication, so you don’t need to movethe sample to and from an ice bath during sonication.
With a programmable sonicator like the EpiShear Probe
Sonicator, you can simply set the duration of the On
and Off pulses, press Start, then walk away. For more
information, visit www.activemotif.com/platform.

Figure 16: The EpiShear Probe Sonicator and the EpiShear Cooled Sonication Platform.
The EpiShear Probe Sonicator is a compact 120-watt unit that can process small or large
samples. The EpiShear Cooled Sonication Platform, sold separately, precisely positions theprobe within each sample, greatly enhancing sample-to-sample reproducibility.

Figure 17: Dounce homogenizer with two pestles.

Whether you shear your chromatinby sonication or enzymatically, thefirst step in preparing your sample
is cell lysis. Active Motif’s Dounce
Homogenizers (Figure 17), which
physically disrupt the cell membranes,
are the best way to prepare lysatesthat are of the highest quality, yieldand concentration.

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