Histone Purification Kit
Histone-Purification-Kit-AM171
Synonyms
| Name | Format | Cat No. | Price |
|---|
Overview
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. The Histone Purification Kit enables you to isolate core histones from any cell culture or tissue sample. The core histones may be purified using the convenient 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 with the efficient gravity flow protocol.
To learn more about the Histone Purification Kit, click on the Description tab below. To view a manual or other related documents, click on the Documents tab. For a complete list of reagents included in the Histone Purification Kit, please select the Contents tab below. Inhibitors are NOT included in the Histone Purification kit and are generally not required to preserve histone post-translational modifications using this kit. If inhibitors are desired, they must be supplied by the user and can be added to the extraction and neutralization buffers during isolation. Additional Extraction Buffer and 5X Neutralization buffer may be purchased separately.
Documents (10)
Description
How does it work?
Unlike histone purification by acid precipitation, our method utilizes a unique purification resin and a series of proprietary elution buffers to isolate very pure histone fractions. The resin has a high binding capacity for histones, so core histones can be isolated from small cell culture samples or from grams of tissue. Sequential elution steps let you collect the histones in either one step containing all core histones or into two separate populations: one enriched for H2A and H2B and a second fraction containing >90% pure H3 and H4 (Figure 1).
Figure 1: SDS-PAGE analysis of histone fractions.Ten µg of sample were loaded per lane. Left panel: Core histones purified from logarithmically growing (Log.), mitotic arrested and sodium butyrate-treated HeLa cells. Center panel: Core histones isolated from rat brain tissue. Right panel: Separate H2A/H2B and H3/H4 fractions from HeLa cells.
Histone Purification Kits include a reusable purification column and neutralization, equilibration, wash and H2A/H2B & H3/H4 elution buffers so you can easily purify core histones from your tissue or cell sample. The binding capacity of the purification resin is robust, so histones can be purified from as little as a single 100 mm plate, or from liters of culture or grams of tissue. The yield of core histones is about 0.4-0.5 mg of total core histones per 3 x 150 mm plates or 2.5 mg per gram of tissue, and the purified histones are ready for downstream analysis.
Core histones isolated by the Histone Purification Kit are highly pure and suitable substrates for downstream assays. The purified histones can also be used with our Chromatin Assembly Kit to enable the generation of chromatin that very closely resembles native chromatin for functional assays.
Advantages
- Separates H2A/H2B histones from H3/H4 histones
- Preserves modifications on histones
- Novel purification resin and elution system yields highly pure histones
Contents
Contents & Storage
Please note that the Histone Purification Kit is shipped at room temperature and contains reagents with multiple storage temperatures inside. Please store each component at the temperature indicated below. All reagents are guaranteed stable for 6 months from date of receipt when stored properly. Each Histone Purification Kit supplies sufficient reagents to perform 10 purifications. This kit includes the following components:
- Extraction Buffer; Store at 4°C
- 5X Neutralization Buffer; Store at 4°C
- Equilibration Buffer; Store at 4°C
- Histone Wash Buffer; Store at 4°C
- Histone H2A/H2B Elution Buffer; Store at 4°C
- Histone H3/H4 Elution Buffer; Store at 4°C
- Purification Resin; Store at 4°C
- Purification Column; Store at RT prior to assembly, then store at 4°C