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   Home  »  Histone Modification Studies 
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Complete Solutions for Histone Modification Studies

We offer the most comprehensive selection of Histone Modification research products to cover every step of the experiment workflow, from upstream to downstream.

Click on any of the steps below to jump
to the respective section

Histone Modification Studies

Histones are primary protein components of eukaryotic chromatin and play a role in gene regulation. H3 and H4 histones have tails protruding from the nucleosome that can be modified post-translationally to alter the histone's interactions with DNA and nuclear proteins, leading to epigenetic changes for regulating many normal and disease-related processes. EpiGentek offers a complete series of kits available for the quantification of methylation, acetylation, and phosphorylation of H3 histones at all sites (see our informative histone modification table).

Histone Methylation

The mechanism known as histone methylation is a post-translational epigenetic modification that involves the transfer of methyl groups to histone proteins via histone methyltransferases (HMTs). Methyl groups are added to the “tails” that protrude from the histone proteins, which is the most common location for post-translational modifications, especially N-terminal tails. Alternatively, histone demethylation is the removal of methyl groups from the histone tails catalyzed by histone demethylases (HDMs). Histone methylation and histone demethylation are epigenetic modifications that have the power to reduce or bolster gene expression, especially as a result of altering chromatin structure.

A histone is a protein that helps to comprise the structure of chromatin, which is composed of DNA-wrapped protein octamers. These octamers consist of duplicates of four core histones (H2A, H2B, H3, and H4). This unit of chromatin is commonly known as a nucleosome. Transcriptional repression or activation can occur as a result of histone methylation or demethylation due to the loosening or restriction of the chromatin structure. Heterochromatin and euchromatin refer to chromatin structures that are either tightly or loosely packed DNA around histones, respectively. Heterochromatin is transcriptionally inactive, whereas euchromatin is transcriptionally active.

Where and how many methyl groups are added to the histones largely determines whether the chromatin is available for transcription or not. Tail residues — lysine (K) and arginine (R) — may be methylated at varying degrees with differing outcomes. For instance, when histone H4 is monomethylated on lysine 20 (H4K20me1), this common histone modification results in the contraction of chromatin. Restricting the chromatin structure prevents transcription from occurring and reduces gene expression. Alternatively, monomethylation of histone H3 on arginine 17 (H3R17me1) leads to transcriptional activation.

Lysine methylation is involved in both transcriptional activation (H3K4, K36, K79) as well as silencing (H3K9, K27, H4K20). Thus, studying the effects of these modifications will allow researchers to better understand how the transfer or removal of different amounts of methyl groups to or from various lysine or arginine residues will impact gene expression. The “histone code” attempts to describe the way histone modifications function together in various combinations to control certain cellular processes.

By measuring histone modifications, researchers can uncover novel epigenetic insight into cellular processes and disease states. Abnormal modifications, for example, have been connected to numerous different diseases, ranging from cancer to autoimmune disorders and inflammatory and neurological diseases. In addition to better understanding the epigenetic underpinnings of the pathological disease process, detecting histone modifications can also aid in the development of histone modification-targeted drugs.

Histone Acetylation

Histone acetylation is an epigenetic modification characterized by the addition of an acetyl group to histone proteins, specifically to the lysine residues within the N-terminal tail. This histone modification is catalyzed by enzymes known as histone acetyltransferases (HATs). The two different types of HATs – cytoplasmic and nuclear – are determined based on intracellular location and histone specificity. Alternatively, histone deacetylases (HDACs) act to remove acetyl groups in a process known as histone deacetylation.

Similar to other histone modifications, histone acetylation/deacetylation impacts chromatin structure and, in turn, gene expression by making the DNA more or less accessible to transcription. Acetylation of lysine residues leads to a transcriptionally active chromatin structure (euchromatin), and deacetylation leads to an inactive, condensed chromatin structure (heterochromatin).

There are four major classes of HDACs: Class I, Class II, Class III, and Class IV. HDAC1, HDAC2, HDAC3 and HDAC8 belong to Class I. HDAC4, HDAC5, HDAC6, HDAC7, HDAC9 and HDAC10 belong to Class II. Seven sirtuins, including SIRT1 through SIRT7, belong to Class III. Lastly, Class IV consists of only HDAC11. These classifications are based on their homology to yeast proteins.

By studying histone acetylation and deacetylation, researchers can gain more insight into the “histone code.” This research can also help contribute to the development of HDAC-targeted drugs. For instance, histone deacetylase inhibitors (HDACi) are often used as mood stabilizers and anti-epileptics. They have also been applied as possible cancer neurodegenerative and inflammatory disease treatment. Interestingly, HDAC inhibitors are known to have specificity toward tumor cells, explaining their widespread use as anticancer drugs.

Histone Phosphorylation

Histone phosphorylation is a post-translational modification that affects serine, threonine, and tyrosine residues. It involves the addition of a phosphoryl group to histone tails, which can play a part in chromatin remodeling. It is possible for all four nucleosomal histone tails to be phosphorylated. One of the best-known functions of histone phosphorylation involves the cellular response to DNA damage. Although numerous phosphorylation sites have been discovered, new sites are being uncovered in ongoing epigenetic research.


Histone Studies Step Details


Histone Extraction

The EpiQuik Total Histone Extraction Kits allow the researcher to reliably and conveniently isolate histone proteins from mammalian cells or tissues in just one hour. These kits are recommended for preparing histone extracts for use in EpiGentek’s histone modification assay kits or a variety of downstream applications for histone acetylation, methylation, phosphorylation, and sumoylation.

Catalog #
Name
Description
OP-0006
EpiQuik Total Histone Extraction Kit
For isolating histone proteins with post-translational modifications (PTMs) intact
OP-0007
EpiQuik Total Histone Extraction HT Kit
For isolating histone proteins from mammalian cells cultured on a 96-well plate


Nuclear Extraction

The EpiQuik Nuclear Extraction Kits allow the researcher to reliably and conveniently isolate nuclear proteins from cells or tissues in just one hour. The extracts can be used in EpiGentek’s DNMT, HMT, Histone Demethylase, HDAC, & HAT kits or in a variety of applications, including Western blots, protein-DNA binding assays, nuclear enzyme assays, and any procedure requiring optimized nuclear proteins.

Catalog #
Name
Description
OP-0002
EpiQuik Nuclear Extraction Kit
For the isolation and extraction of nuclear proteins from cells or tissues while keeping enzymatic activity intact
OP-0022
EpiQuik Nuclear Extraction Kit II (Nucleic Acid-Free)
For isolation of nucleic acid-free nuclear proteins from cells or tissues while keeping enzymatic activity intact


Histone H3 & H4 Multiplex Assays

With the EpiQuik Histone H3 and H4 Modification Multiplex Assay Kits, you can get an overall view of your sample’s histone modification status. This ELISA-like method requires only a standard microplate reader. Save time and money by simultaneously measuring 21 different histone H3 modifications, 10 different histone H4 modifications, or 22 different circulating histone H3 modifications. Each of these kits includes all of the most important and well-characterized patterns.

Catalog #
Name
Description
P-3100
EpiQuik Histone H3 Modification Multiplex Assay Kit
For ELISA-like quantitation of 21 different histone H3 modification patterns on the same plate
P-3102
EpiQuik Histone H4 Modification Multiplex Assay Kit
For ELISA-like quantitation of 10 different histone H4 modifications on the same plate
P-3106
EpiQuik Circulating Modified Histone H3 Multiplex Assay Kit
For ELISA-like quantitation of 22 different circulating histone H3 modifications on the same plate


Histone Quantification

Quantifying histone methylation or acetylation is particularly useful for studying gene expression patterns. EpiGentek offers a complete series of ELISA-like kits to quantify H3 or H4 methylation or acetylation at specific lysines in both colorimetric and fluorometric versions.

Histone H3 and H4 Methylation Quantification Kits Include: (*Fluorometric Version)

Histone Cat# Histone Cat# Histone Cat# Histone Cat#
H3K4me1 P-3024 H3K9me3 P-3034 H3K36me1 P-3046 H3K79me3 P-3058
H3K4me1* P-3025 H3K9me3* P-3035 H3K36me1* P-3047 H3K79me3* P-3059
H3K4me2 P-3022 H3K9Pan P-3036 H3K36me2 P-3048 H3K79Pan P-3060
H3K4me2* P-3023 H3K9Pan* P-3037 H3K36me2* P-3049 H3K79Pan* P-3061
H3K4me3 P-3026 H3K27me1 P-3038 H3K36me3 P-3050 H4K20me1 P-3064
H3K4me3* P-3027 H3K27me1* P-3039 H3K36me3* P-3051 H4K20me1* P-3065
H3K4Pan P-3028 H3K27me2 P-3040 H3K36Pan P-3052 H4K20me2 P-3066
H3K4Pan* P-3029 H3K27me2* P-3041 H3K36Pan* P-3053 H4K20me2* P-3067
H3K9me1 P-3030 H3K27me3 P-3042 H3K79me1 P-3054 H4K20me3 P-3068
H3K9me1* P-3031 H3K27me3* P-3043 H3K79me1* P-3055 H4K20me3* P-3069
H3K9me2 P-3032 H3K27Pan P-3044 H3K79me2 P-3056 H4K20Pan P-3070
H3K9me2* P-3033 H3K27Pan* P-3045 H3K79me2* P-3057 H4K20Pan* P-3071

Histone H3 and H4 Acetylation Quantification Kits Include: (*Fluorometric Version)

Histone Cat# Histone Cat# Histone Cat# Histone Cat#
H3K9ac P-4010 H3K18ac* P-4015 H3K56ac* P-4021 H4K12ac P-4028
H3K9ac* P-4011 H3K27ac P-4059 H4K5ac P-4022 H4K12ac* P-4029
H3K14ac P-4012 H3K36ac P-4018 H4K5ac* P-4023 H4K16ac P-4026
H3K14ac* P-4013 H3K36ac* P-4019 H4K8ac P-4024 H4K16ac* P-4027
H3K18ac P-4014 H3K56ac P-4020 H4K8ac* P-4025    

View more Histone H3 and H4 Methylation or Acetylation Quantification Kits.


Histone Acetyltransferase (HAT) and Histone Deacetylase (HDAC)

The balance between protein acetylation and deacetylation controls several physiological and pathological cellular processes. Study the enzymes involved in maintaining this equilibrium - histone acetyltransferases (HATs) and histone deacetylases (HDACs). We have kits that can quickly measure HAT activity or quantify any of the entire family of HDACs (1-11).

Catalog #
Name
Description
P-4003
EpiQuik HAT Activity/Inhibition Assay Kit
Measure histone acetyltransferase activity/ inhibition safely and quickly
P-4034
P-4035*
Epigenase HDAC Activity/Inhibition Direct Assay Kits (*Fluorometric Version)
Measure total activity/inhibition of HDAC enzymes in nuclear extracts or purified HDAC isoforms (HDACs 1-11)
See Cat#s
EpiQuik HDAC1 -7 Assay Kits
Measure the total amount of HDAC 1-11 proteins quantitatively through an ELISA-like method

Histone Methyltransferase (HMT) & Demethylase

Lysine methylations mark various locations on the histone tail and globular domains, and their levels are specifically balanced by methyltransferases (‘writers’) and demethylases (‘erasers’). Study the function of histone methyltransferase (HMT) or demethylase using our convenient ELISA-like assays that directly measure HMT or histone demethylase activity level of your nuclear extract of purified enzyme samples.

Histone Methyltransferase (HMT) Assays Include:

Cat #
Name
Description
P-3002
EpiQuik Histone Methyltransferase Activity/Inhibition Assay Kit (H3K4)
Measure activity of HMTs that specifically target histone H3 at lysine 4
P-3003
EpiQuik Histone Methyltransferase Activity/Inhibition Assay Kit (H3K9)
Measure activity of HMTs that specifically target histone H3 at lysine 9
P-3005
EpiQuik Histone Methyltransferase Activity/Inhibition Assay Kit (H3K27)
Measure activity of HMTs that specifically target histone H3 at lysine 27
P-3088
Epigenase PRMT Methyltransferase (Type II-Specific) Activity/Inhibition Assay Kit
Measure activity of total type II PRMT using nuclear extracts or purified enzymes

Histone Demethylase Assays Include: (*Fluorometric Version)

Cat #
Name
Description
P-3078
P-3079*
Epigenase LSD1 Demethylase Activity/Inhibition Assay Kits
Measures LSD1 activity via a straightforward detection of LSD1-converted demethylated products
P-3080
P-3081*
Epigenase JMJD2 Demethylase Activity/Inhibition Assay Kits
Measure JMJD2 activity via a straightforward detection of JMJD2-converted demethylated products
P-3082
P-3083*
Epigenase JARID Demethylase Activity/Inhibition Assay Kits
Measure JARID activity via a straightforward detection of JARID-converted demethylated products
P-3084
P-3085*
Epigenase JMJD3/UTX Demethylase Activity/Inhibition Assay Kits
Measure JMJD3 or UTX activity via a straightforward detection of JMJD3/UTX-converted demethylated products



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