Histone Purification Microplate Kit

by Active Motif
Available SKUs: 40027 |
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Description

Active Motif's Histone Purification Microplate Kit enables purification of core histones (H2A, H2B, H3 and H4) using very small amounts (as little as 0.9 x 105 cells) of starting material, and its microplate spin column format enables elution into a 96-well plate to easily transition into downstream high-throughput applications.

PBMC ChIP on CD8+ T Cells
Achieve better yields with Histone Microplate Purification than with crude extraction.

(Click image to enlarge)

The Histone Purification Microplate Kit method utilizes the identical proprietary buffer system used in our Histone Purification Kit and our Histone Purification Mini Kit to purify core histones from cells and tissue samples. This purification method is superior to acid precipitation methods because it preserves post-translational histone modifications and greatly improves the purity and yield of your isolated histones to improve the reliability and consistency of your sample preparation. Histones isolated by this method are excellent substrates for use in downstream high-throughput screening assays, Western blot or mass spectrometry.

Advantages

  • Only commercial kits available for histone purification
  • Higher quality sample prep than crude acid extraction
  • 96-well format with modular stripwell design for flexibility in throughput
  • Preserves modifications on histones
  • Purify histones from as few as 0.9x105 cells
  • Works with cell or tissue samples

To learn more about the Histone Purification Microplate Kit, click on the Method or Data, or tabs 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 Microplate Kit, please select the Contents tab below. Inhibitors are NOT included in the Histone Purification Microplate 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.

 
Name Format Cat No. Price  
Histone Purification Microplate Kit 96 rxns 40027 $820 Buy Now
Protease Inhibitor Cocktail 1 ml 37490 $125 Buy Now
Protease Inhibitor Cocktail 5 ml 37491 $360 Buy Now
Phosphatase Inhibitor Cocktail 1 ml 37492 $165 Buy Now
Phosphatase Inhibitor Cocktail 5 ml 37493 $530 Buy Now
Deacetylase Inhibitor 5 ml 37494 $160 Buy Now
Extraction Buffer 100 ml 37513 $65 Buy Now
5X Neutralization Buffer 50 ml 37514 $65 Buy Now

How does it work?

The Histone Purification Microplate Kit is designed to provide researchers with a rapid method for purification of core histones to streamline into downstream high throughput screening applications. The method improves yield, purity and concentrations above what is achievable with crude acid extraction, providing the highest quality substrate for use in biopharmaceutical and screening assays.

The kit supplies enough reagents to perform up to 96 histone purifications starting with as little as a 0.9x105 cells. The method utilizes column filtration to simplify and speed up the processing of samples. The columns are provided in a 96-well plate spin column format that features a modular 8-strip unit design to allow adaptation to both low and high throughput processing.

Histone Purification Microplate

The Histone Purification Microplate Kit utilizes a series of proprietary elution buffers that are identical to those used in our Histone Purification Kit and our Histone Purification Mini Kit to purify core histones from cells and tissue samples. Purification of histones using this method eliminates contaminating acid-insoluble cellular proteins that are left behind with standard acid extraction protocols. Removal of these impurities also prevents any residual alterations to the histone modifications so that key post-translational modifications on histone tails are preserved, providing you with the highest quality histone samples for use in your downstream analysis.

The kit method is simple. First, make an extract from your cell or tissue samples. Then, apply the extract to the purification, elute histones and quantititate. Core histones (H2A, H2B, H3 and H4) are eluted as one highly concentrated total population.

Quantification of histone proteins

Quantification of histone proteins from small cell numbers is not possible using traditional methods for protein quantification, such as Bradford or absorbance at 230 nm, as the yield will fall below the detectable range for these methods. Instead, we suggest using Active Motif’s Histone H3 Total ELISA Kit to quantify the amount of histone H3 recovered. ELISAs for specific histone modifications are also available to quantify the amount of a particular histone PTM within the sample.

  • Histone H3 Total ELISA Kit (Cat. No. 53110) – we suggest using a starting volume of 2 µl per well of purified histones extracted from 1 x 105 cells in the ELISA.
  • Histone Modification ELISAs – we suggest using a starting volume of 20 µl per well of purified histones extracted from 1 x 105 cells in the ELISA.

Achieve the Highest Quality Samples for Your High Throughput Applications

The Histone Purification Microplate Kit provides a fast, reliable and high throughput method for for obtaining high quality histone samples for use in downstream applications. The proprietary method enables purification of core histones at significantly higher yields and purity than what is achievable by crude extraction.

Purity of the core histones isolated with the Histone Purification Microplate Kit
Figure 1: SDS-PAGE comparison of the purity of histone fractions isolated by crude acid extraction or purified using the Histone Purification Microplate Kit.

Histones were extracted from 2 x 105 HeLa cells using either crude acid extraction conditions (Lanes 1-3) or the Histone Purification Microplate Kit (Lanes 4-6). Nine µl of each biological replicate were loaded and run on a 4-20% SDS gradient gel, then stained with Ponceau Stain. The crude acid extracted material shows contamination of other acid soluble proteins in the elution, while the Histone Purification Microplate Kit yields a pure fraction of core histones.

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