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Index / EpigenTek / EpiQuik Total Histone Extraction Kit / Product Detail : OP-0006-100 EpiQuik Total Histone Extraction Kit
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#OP-0006-100 EpiQuik Total Histone Extraction Kit

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  Price : 196   EUR
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222   CHF

Product name : EpiQuik Total Histone Extraction Kit

Catalog number : OP-0006-100

Quantity: 100 extractions

Availability: Yes

Supplier name : EpigenTek

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More Details about

Product Overview

The EpiQuik™ Total Histone Extraction Kit provides a simple and selective method for extracting histone proteins used for a variety of applications, which include histone modification such as acetylation, methylation, and sumoylation. The kit can be used to extract histones from mammalian cells and tissues with the fastest procedure available on the current market, which can be finished within 60 minutes. The EpiQuik™ Total Histone Extraction Kits are also specifically designed to meet the requirements of histone extracts used in EpiQuik™ assays. Yield of the total histone proteins is approximately 0.4 mg per 107 cells or 100 mg of tissue. The yield may vary depending on cell or tissue type.

 

Product Details

The EpiQuik™ Total Histone Extraction Kit simply applies our proprietary histone isolation buffers to cells/tissues. After treatment with pre-lysis, lysis, and balance buffers, the total histones are easily extracted for immediate use or storage at proper conditions.

SCHEMATIC PROCEDURE:  



Histone extracts were prepared from MCF-7 cells using the EpiQuik Total Histone Extraction Kit and acetyl histone H3-K9 was quantified using the EpiQuik Global Acetylated Histone H3-K9 Quantification Kit (Fluorometric) (Cat. No. P-4011).
Product Components

10X Pre-Lysis Buffer
Lysis Buffer
Balance Buffer
DTT Solution
User Guide

 

Frequently Asked Q's

1. Will the buffer affect the histone modification status such as acetylation or methylation?
No, it will not.

2. What is the minimal amount of tissue that can be used for extraction?
1 mg, but for the best results, we recommend more than 10 mg.

 

Product Citations

Yamamura, Y. et. al. (October 2011). TNF-a inhibits aquaporin 5 expression in human salivary gland acinar cells via suppression of histone H4 acetylation. J Cell Mol Med. 141(8):1464-8. PubMed Abstract

Jing, L. et. al. (April 2011). Effect of the histone deacetylase inhibitors on behavioural sensitization to a single morphine exposure in mice. Neurosci Lett. 494(2): 169-73. PubMed Abstract 

Kawakami, K. et. al. (January 2011). Long interspersed nuclear element-1 hypomethylation is a potential biomarker for the prediction of response to oral fluoropyrimidines in microsatellite stable and CpG island methylator phenotype-negative colorectal cancer. Cancer Sci. 102(1): 166-74. PubMed Abstract

Hansen, A. et. al. (July 2010). Development of a drug screening platform based on engineered heart tissue. Circulation Research. 107(1): 35-44. PubMed Abstract

Diehl, F. et. al. (March 2010). Cardiac deletion of smyd2 is dispensable for mouse heart development. PLoS One. 5(3): e9748. PubMed Abstract

 

Shipping and Storage
The EpiQuik™ Total Histone Extraction Kit is shipped at ambient room
temperature.
Upon receipt: (1) Store DTT Solution at 4°C; and (2) Store all remaining
components at room temperature.
All components of the kit are stable for 6 months from the date of
shipment, when stored properly.

 

Introduction
Product Use Information
Uses The EpiQuik™ Total Histone Extraction Kit is suitable
for a quick preparation of total histone extracts from
mammalian cells and tissue samples.
Input
Amount
The minimal amount of starting materials can be as
low as 105 cells or 1 mg tissue. For the best results,
the cell number should be greater than 106 cells or the
tissue amount should be greater than 10 mg. A total
of 100 standard extractions (use 107 cells or 100 mg
tissues per extraction) can be performed with this kit.
Yield Yield of the total histone proteins can be up to 0.4 mg
per 107 cells or per 100 mg tissues. The yield may
vary depending on the cell or tissue type.
Precautions To avoid cross-contamination, carefully pipette the
sample or solution into the strip wells. Use aerosolbarrier
pipette tips and always change pipette tips
between liquid transfers. Wear gloves throughout the
entire procedure. In case of contact between gloves
and sample, change gloves immediately.

General Product Information

Quality Control
Each lot of the EpiQuik™ Total Histone Extraction Kit
is tested against predetermined specifications to
ensure consistent product quality. Epigentek
guarantees the performance of all products in the
manner described in our product instructions.

Product Warranty
If this product does not meet your expectations,
simply contact our technical support unit or your
regional distributor. We also encourage you to contact
us if you have any suggestions about product
performance or new applications and techniques.
Safety Suitable lab coat, disposable gloves, and proper eye
protection are required when working with this
product.

Product Updates
Epigentek reserves the right to change or modify any
product to enhance its performance and design. The
information in this User Guide is subject to change at
any time without notice. Thus, only use the User
Guide that was supplied with the kit when using that
kit.

Usage Limitation
The EpiQuik™ Total Histone Extraction Kit is for
research use only and is not intended for diagnostic or
therapeutic application.

Intellectual Property
The EpiQuik™ Total Histone Extraction Kit and
methods of use contain proprietary technologies by
Epigentek.

A Brief Overview
Histones are the chief protein components of chromatin in biology.
They act as spools around which DNA winds, and also play a role in
gene regulation.
The core histones include H2A, H2B, H3, and H4. Histones undergo
posttranslational modifications, which alter their interaction with DNA
and nuclear proteins. The H3 and H4 histones have long tails
protruding from the nucleosome, which can be covalently modified at
several places. Modifications of the tail include methylation,
acetylation, phosphorylation, ubiquitination, sumoylation, citrullination,
and ADP-ribosylation (H2A can also be modified). Combinations of
modifications are thought to constitute a code, the so-called "histone
code." Histone modifications act in diverse biological processes such
as gene regulation, DNA repair and chromosome condensation
(mitosis).
The EpiQuik™ Total Histone Extraction Kit provides a simple and
selective method for extracting histone proteins used for a variety of
applications, which include histone modifications such as acetylation,
methylation, and sumoylation. The EpiQuik™ Total Histone
Extraction Kit is also specifically designed to meet the requirements
of histone extracts used in EpiQuik™ histone quantification assays.
The EpiQuik™ Total Histone Extraction Kit can be used to extract
histones from mammalian cells and tissues. The EpiQuik™ Total
Histone Extraction Kit has the fastest procedure available on the
market, allowing completion within 60 minutes.

Usage
Protocol
For the best results, please read the protocol in its entirety prior to
starting your experiment.

1.
For Tissues
(Treated and Untreated)
a. Weigh the sample and cut the sample into
small pieces (1-2 mm3) with a scalpel or
scissors.
b. Transfer tissue pieces to a Dounce
homogenizer.
c. Dilute 10X Pre-Lysis Buffer into 1X Pre-Lysis
Buffer with distilled water at a 1:10 ratio (e.g., 1
ml of 10X Pre-Lysis Buffer + 9 ml of water).
d. Add the Diluted 1X Pre-Lysis Buffer at 200
mg/ml, and disaggregate tissue pieces by 50-
60 strokes.
e. Transfer homogenized mixture to a 15 ml
conical tube and centrifuge at 3000 rpm for 5
min at 4°C. If total mixture volume is less than 2
ml, transfer mixture to a 2 ml vial and centrifuge
at 10,000 rpm for 1 min at 4°C.
f. Remove supernatant.

1.
For Cells
(Treated and Untreated)
a. Harvest cells and pellet the cells by
centrifugation at 1000 rpm for 5 min at 4°C.
b. Dilute 10X Pre-Lysis Buffer into 1X Pre-Lysis
Buffer with distilled water at a 1:10 ratio (e.g., 1
ml of 10X Pre-Lysis Buffer + 9 ml of water).
c. Re-suspend cells in the Diluted 1X Pre-Lysis
Buffer at 107 cells/ml and lyse cells on ice for
10 min with gentle stirring.
d. Centrifuge at 3000 rpm for 5 min at 4°C. If cell
lysates are prepared in a 1.5 to 2 ml size vial,
centrifuge at 10,000 rpm for 1 min at 4°C.
e. Remove supernatant.
2. a. Re-suspend cell/tissue pellet in 3 volumes
(approximately 200 ,l/107 cells or 100 mg
tissues) of Lysis Buffer and incubate on ice for
30 min.
3.
a. Centrifuge at 12,000 rpm for 5 min at 4°C and
transfer the supernatant fraction (containing
acid-soluble proteins) into a new vial.
4.
a. Prepare Balance-DTT Buffer by adding DTT
Solution to Balance Buffer at a 1:500 ratio
(e.g., 1 ,l of DTT Solution + 500 ,l of Balance
Buffer).
b. Add 0.3 volumes of the Balance-DTT Buffer to
the supernatant immediately (e.g., 0.3 ml of
Balance-DTT Buffer to 1 ml of supernatant).
5.
a. Quantify the protein concentration with an OD
reading. BSA can be used as a standard.
6. a. Aliquot and store the extract at –20°C for
several days, or –80°C for long-term storage.
Avoid repeated thawing and freezing.

Note: If salt precipitates are seen in the extracts
after being frozen, warm the extracts at room
temperature for several minutes and pipette around
several times until salts are re-dissolved.

 

Related Products
Protein Preparation
Base Cat. # Description
OP-0002 EpiQuik™ Nuclear Extraction Kit
OP-0022 EpiQuik™ Nuclear Extraction Kit II (Nucleic Acid-Free)
OP-0003 EpiQuik™ Whole Cell Extraction Kit

 

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