Core Histone Purificationwww.activemotif.com

Histone Purification Kit
purify core histones while preserving modification states

Active Motif’s Histone Purification Kit enables you to purify core histone proteins from cell or tissue samples while
preserving post-translational modifications such as acetylation, methylation, and phosphorylation states (Figure 1).
This method utilizes purification resin and a proprietary buffer system to purify the core histones. Unlike standard
acid extraction methods, this kit enables you to further separate the total core histones into two populations: one
fraction enriched for H2A and H2B dimers and a highly pure fraction containing H3 and H4 tetramers (Figure 2).
Separate H2A/H2B from H3/H4Histone Purification Kits provide you with areusable purification column, Neutralization,Equilibration & Wash Buffers plus separate
H2A/H2B and H3/H4 Elution Buffers so you
can easily purify core histones from tissue
or cell samples. The binding capacity of thepurification resin is robust, so histones can bepurified from as little as one 100 mm plate orliters of culture or grams of tissue. The yieldis ~0.5 mg of total core histones per 3 x 150mm plates, or 2.5 mg from 1 gram of tissue.
Purified histones are ready for downstream
analysis with Active Motif’s many different histone antibodies, as well as with ourChromatin Assembly Kit (Catalog No. 53500);
this unique kit improves functional assays byenabling you to generate chromatin in vitro
that very closely resembles native chromatin.
• Purifies total core histones (H2A, H2B,H3 & H4) or separate fractions of
H2A/H2B dimers and H3/H4 tetramers• Preserves modifications on histones• Novel purification resin and elution
system yields highly pure histones

Active Motif Purification Method
8.0

5X Neutralization Buffer, Extraction Buffer, Equilibration
Buffer, Histone Wash Buffer, H2A/H2B Elution Buffer,
H3/H4 Elution Buffer, Purification Resin and Purification
Column. Reagents are to be stored at 4°C. All reagentsare guaranteed stable for 6 months from date of receipt
when stored properly.

2.0

1.0

Acid Extraction Method

0.5

8.0

4.0

2.0

1.0

0.5µg
Phosphorylation

Acid Extraction Method
SodiumButyrate

Mitotic
Arrested

Active Motif Purification Method
SodiumButyrateLog.

Mitotic
Arrested

Log.
Acetylation

Active Motif Purification Method

Acid Extraction Method
Log.

Mitotic
Arrested

SodiumButyrate

Log.

Mitotic
Arrested

SodiumButyrate

Methylation

Figure 1: Histone Purification Kit preserves post-translational modification states.
Western blot analysis of histone modifications comparing samples isolated by standard acid extraction vs. with theHistone Purification Kit. Top: Histone H3 phospho Ser28 Rat mAb (Clone HTA28) (Cat. No. 39098). Center: Histone
H4 tetra-acetyl Rabbit pAb (Cat. No. 39177). Bottom: Histone H3 trimethyl Lys4 Rabbit pAb (Cat. No. 39159).
MWLog.

MitoticArrested

MWSodium
Butyrate

989864645050363630301664CONTENTS & STORAGE

4.0

MWCore Histones

98645036H3H2B
H2A
H43016H2B
H2A

H3H416H464

H3
H2A/H2B

6Figure 2: Core histones purified from HeLa cells and brain tissue.
10 µg of sample were loaded per lane. Left: Core histones purified from logarithmically growing, mitotic arrested or
sodium butyrate-treated HeLa cells. Center: Core histones isolated from rat brain tissue. Right: Separate H2A/H2Band H3/H4 fractions purified from HeLa cells.

ProductFormat

Catalog No.

Histone Purification Kit

10 rxns40025

North America 877 222 9543 Europe +32 (0)2 653 0001 Japan +81 (0)3 5225 3638