ACTIVE MOTIF

Histone Purification Kits
improve quality, yield and preservation of histone marks during sample prep

Active Motif is the only company to offer a method for purification of core histones. Our Histone Purification Kits utilize aproprietary histone purification technique that preserves histone modifications and produces higher purity and yield than acidextraction methods for higher quality sample preparation. The kits enable purification of core histone proteins from cell or
tissue samples and are available in maxi, mini and microplate-based formats to suit your throughput needs.

Which kit is right for you?

Histone Purification Advantages

Active Motif’s Histone Purification Kits are available in multiple
formats so you can choose the one that best fits the needs ofyour assay. Use Table 1 below to determine which kit is right foryou. For complete details on our histone purification products,
please visit us at www.activemotif.com/histonepur.

• Exclusivity – only commercial kits for histone purification• Quality – enhanced histone purity and preservation ofhistone modifications• Scalability – 3 throughput formats to suit your needs• Flexibility – works with cell or tissue samples

The Histone Purification Method
Active Motif’s Histone Purification, Histone Purification Miniand Histone Purification Microplate Kits are the only commercially available assays for isolating purified core histones. The
assays were designed to overcome the limitations of crude acidextraction methods for isolation of core histones. Our Histone
Purification Kits use a unique purification resin and a series ofproprietary elution buffers to isolate extremely pure fractionsof histones from any cell or tissue sample. Histone purification
eliminates contaminating acid-insoluble cellular proteinsthat are left behind with standard acid extraction protocols.
Removal of these impurities prevents enzymatic alterations tohistone modifications. The results are improved yields, purity,and preservation of modifications above what is achievable withcrude acid extraction (Figure 1) to ensure you have the highest
quality histone samples for use in your downstream analysis.
Kit

Application

Acetylation

Methylation

Phosphorylation

Figure 1: Acetyl, methyl & phosphoryl post-translational modifications are preserved
as well or better with Histone Purification compared to acid precipitation methods.
Western blot analysis of histone modifications comparing samples isolated by standard
acid extraction vs. with the Histone Purification Kit. Top: Histone H3 phospho Ser28 mAb(Cat. No. 39098). Center: Histone H4 tetra-acetyl pAb (Cat. No. 39177). Bottom: Histone
H3 trimethyl Lys4 pAb (Cat. No. 39159).

FormatElutionCapacity

Purifications

Gravity Flow

Separate H2A/H2B &H3/H4 fractions

0.5-2.5 mg

10Spin Column

H2A, H2B, H3 & H4 ina single fraction

0.5-2.5 mg

10Histone Purification
Kit• Low throughput• Large sample quantities

Histone Purification
Mini Kit• Medium throughput• Mid-range samplequantities

Mini Spin Column

H2A, H2B, H3 & H4 ina single fraction

0.1-0.5 mg

20Histone Purification
Microplate Kit• High throughput• Low sample quantities

96 Stripwell Plate
Spin Column

H2A, H2B, H3 & H4 ina single fraction< 0.48 mg

96

Table 1: Specifications for Active Motif’s Histone Purification Kits.

North America 877 222 9543

Europe +32 (0)2 653 0001

Japan +81 (0)3 5225 3638

www.activemotif.com

Total Histone H3 ELISA

3.5
3.02.5
OD 450 nm

Improve the consistency and reliability of your analysis ofhistones and their post-translational modifications by using
purified histone samples. Our proprietary histone purification
method is available in three different format options to suit thethroughput needs of your downstream analysis.

2.0
1.5
1.0Histone Purification Kit

0.5
0
105

2.5

1.25.625 BlankniMintioacxtr ite E te K
udCr roplacMi
Kit

20

AM40

AM

The Histone Purification Kit enables you to isolate core histones from any cell culture or tissue sample. It was designed forlarger sample amounts; the kit contains 10 columns, each with acapacity of 0.5-2.5 mg. Two options are available forpurification – the core histones may be purified using theconvenient spin column format as one total population
containing H2A, H2B, H3 and H4, or further purified into
separate fractions of H2A/H2B dimers and H3/H4 tetramers
by using the efficient gravity flow protocol (Figure 2).

Recombinant H3 protein (ng/well)Figure 3: Active Motif’s Histone Purification Mini and Microplate kits show improved
histone yield over crude acid extraction as measured using the Total Histone H3 ELISA Kit.
The Total Histone H3 ELISA Kit (Catalog No. 53110) was used to generate a standard curvewith the kit’s Recombinant Histone H3 protein (red). The kit was then used to quantify
the amount of Total H3 in purified histone samples obtained using the Histone Purification
Mini Kit and the Histone Purification Microplate Kit as compared to samples isolatedusing crude acid extraction. Results show Active Motif’s histone purification method
significantly improves yield over crude acid extraction.

Histone Purification Microplate Kit

Figure 2: SDS-PAGE of histone fractions purified using the Histone Purification Kit.
10 µg of sample were loaded per lane. Left panel: single fractions of core histones purified from logarithmically growing tissue culture cells (TC) and core histones isolated fromrat brain tissue (Brain). Right panel: separate H2A/H2B and H3/H4 fractions purified fromHeLa cells.

The Histone Purification Microplate Kit offers the same
proprietary purification protocol as the original Histone Purification Kit, but uses a convenient plate-based format for highthroughput processing of small sample volumes. This kit is idealfor researchers with limited starting material or who are seeking
a method to readily transition their samples to downstream
96-well plate-based screening or profiling assays, such as ourHistone Modification ELISAs (Figure 3). Similar to the Mini Kit,
the Histone Purification Microplate Kit purifies core histones as
one total (H2A, H2B, H3 and H4) population; it cannot fractionate the histones into discrete H2A/H2B and H3/H4 fractions.
To learn more about our products for histone purification,please visit us at www.activemotif.com/histonepur.

Histone Purification Mini Kit
The Histone Purification Mini Kit offers the same proprietary
purification protocol as the original Histone Purification Kit
but was designed to process smaller samples. It uses a moreconvenient mini spin column-based format, each with a capacity of 0.1-0.5 mg. This kit enables 20 purifications and allows
for extraction of the core histones as one total population.
However, unlike the original kit, the Mini Kit cannot be used toseparate histones into discrete H2A/H2B and H3/H4 fractions.

North America 877 222 9543

Europe +32 (0)2 653 0001

ProductFormat

Cat. No.

Histone Purification Kit

10 rxns

40025

Histone Purification Mini Kit

20 rxns

40026

Histone Purification Microplate Kit

96 rxns

40027

TO PLACE AN ORDER, call us or send an email to orders@activemotif.com.

Japan +81 (0)3 5225 3638

www.activemotif.com