
Mod Spec® は、質量分析計を使って60種類以上の異なるヒストン修飾の相対量を一度に解析する手法です. Mod Spec® は、エピジェネティックな変化が疾患や薬物処理に応答して生じているかどうかを判断するための出発点になります。多くの修飾変化を一度に見出すことで、予想された変化の確認だけでなく、予期しない重要な変化などを発見できることも期待できます。
日本語チラシはこちら。
ヒストン修飾変化の解析例:
- エピジェネティックな阻害剤への反応
- 正常細胞と罹患細胞の比較
- Knock-out 細胞 や動物細胞
- 薬剤処理した移植サンプル
- ヒト生検サンプル
What our customers are saying about us...
"While working on the molecular mechanism of an epigenetic drug, we outsourced Mod Spec® to Active Motif in order to get a broader overview of the drug-induced changes of histone modifications. Overall, we were very pleased with the quality of the service, the kept timeline and last but not least, the fair price. We found surprising things that were not on our radar before."
Matthias Lauth, PhD
Phillips University
Marburg, Germany
View complete list of testimonials >
プロセス
細胞ペレットや組織をActive Motif にお送りいただければ、ヒストンの抽出、サンプル調製を行い、Thermo Scientific™ TSQ Quantum Ultra™ Triple-Stage Quadrupole Mass Spectrometerにて分析を行います. すべてのサンプルは、トリプリケートで解析し、理解しやすい形のデータに変換してお渡しいたします。通常は、サンプルを受領後4-6週間で結果をお知らせいたします。
必要なサンプル量
- 細胞: 2 - 5 million
- 組織: 25 - 100 mg
Mod Spec® 受託解析の見積もり依頼Services Inquiry
Histone modifications detected by Mod Spec® | ||
---|---|---|
H1.4: K25UN | H1.4: K25AC | H1.4: K25ME1 |
H1.4: K25ME2 | H1.4: K25ME3 | ^H2A: K5UN |
^H2A: K5AC | ^H2A: K9UN | ^H2A: K9AC |
^H2A: K36UN | ^H2A: K36AC | ^H2A1: K13UN |
^H2A1: K13AC | ^H2A1: K15UN | ^H2A1: K15AC |
^H2A1: K15UB | ^H2A3: K13UN | ^H2A3: K13AC |
^H2A3: K15UN | ^H2A3: K15AC | ^H2A3: K15UB |
*H3R2UN: K4UN | *H3R2UN: K4AC | *H3R2UN: K4ME1 |
*H3R2UN: K4ME2 | *H3R2UN: K4ME3 | H3R2UN: Q5UN |
H3R2UN: Q5ME1 | H3: K9UN | H3: K9AC |
H3: K9ME1 | H3: K9ME2 | H3: K9ME3 |
H3: K14UN | H3: K14AC | H3: K18UN |
H3: K18AC | H3: K18ME1 | H3: Q19UN |
H3: Q19ME1 | H3: K23UN | H3: K23AC |
H3: K23ME1 | H3: R42UN | H3: R42ME2 |
H3: R49UN | H3: R49ME2 | H3: Q55UN |
H3: Q55ME1 | H3: K56UN | H3: K56AC |
H3: K56ME1 | H3: K64UN | H3: K64AC |
H3: K79UN | H3: K79AC | H3: K79ME1 |
H3: K79ME2 | H3: K79ME3 | H3: K122UN |
H3: K122AC | ♦H3.1: K27UN | ♦H3.1: K27AC |
♦H3.1: K27ME1 | ♦H3.1: K27ME2 | ♦H3.1: K27ME3 |
♦H3.1: K36UN | ♦H3.1: K36AC | ♦H3.1: K36ME1 |
♦H3.1: K36ME2 | ♦H3.1: K36ME3 | H3.3: K27UN |
H3.3: K27AC | H3.3: K27M | H3.3: K27ME1 |
H3.3: K27ME2 | H3.3: K27ME3 | H3.3: K36UN |
H3.3: K36AC | H3.3: K36ME1 | H3.3: K36ME2 |
H3.3: K36ME3 | H4: K5UN | H4: K5AC |
H4: K8UN | H4: K8AC | H4: K12UN |
H4: K12AC | H4: K16UN | H4: K16AC |
H4: K20UN | H4: K20AC | H4: K20ME1 |
H4: K20ME2 | H4: K20ME3 |
^ Multiple H2A isoforms may contribute to the signal for modifications on H2A.
* H3R2me2 and H3K4me2/3 are mutually exclusive modifications. H3R2 methylation prevents H3K4 methylation. Therefore, H3K4 modifications are reported only on the H3R2 unmodified peptide. For more information, see Nature. 2007 Oct 18; 449(7164):933-7.
♦ H3.2 may contribute to signals for modifications labeled H3.1.
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- Mod Spec® Service Data
- How does Mod Spec® work?
- Mod Spec® Services Publications
Name | Cat No. | Price | |
---|---|---|---|
Mod Spec® | 25085 | Get Quote |
Figure 1: Mod Spec® data from control HeLa cells and HeLa cells treated for 7 days with 0.5uM GSK-126.(Click image to enlarge)
GSK126 is an inhibitor that blocks the methytransferase activity of EZH2, resulting in global decreases in H3K27me2 and H3K27me3. A) A selection of H3 modifications are shown from Active Motif’s Mod Spec® assay that confirm significant decreases in H3K27 methylation with concomitant increases in acetylated and unmodified H3K27 (highlighted in yellow). Changes at H3K36 were also detected (highlighted in red). B) A selection of H4 modifications do not show significant changes in response to GSK126 treatment.
Figure 2: Mod Spec® data from control HEK293 cells and HEK293 cells treated for 6 hours with 5 mM sodium butyrate (NaB).
NaB is a general HDAC inhibitor and treatment is expected to increase histone acetylation levels. A) Mod Spec® data for a selection of acetylated and unmodified histone sites is shown. Acetylation is increased at all sites after treatment. Unmodified peptide is displayed for some sites and shows a concomitant decrease after NaB treatment. B) Methylation at various histone residues is not affected by NaB treatment.
How does Mod Spec® work?
- Histones are extracted from samples.
- Derivatization of histones using propionic anhydride.
- Mass spectrometry of proteins requires digestion of the proteins into small peptides before being injected into the mass spectrometer. However, if peptides are too small they can not be separated in the LC step and if too big they are too complex to analyze. The most common method of digestion is with trypsin, an enzyme that cleaves proteins at lysine and arginine residues. This presents a problem when analyzing histones since histone tails are rich in lysine, resulting in digested peptides that are too small. Propionylation, using propionic anhydride, blocks cleavage at lysines while allowing digestion at arginines to occur. The resulting peptides are of the appropriate size for mass spec analysis.
- Histones are digested with trypsin.
- Samples are analyzed on a Triple-Stage Quadrupole Mass Spectrometer.
- Peptides of the appropriate molecular weight are selected.
- Peptides are further fragmented in the collision chamber.
- Peptides fragments of interest are selected and analyzed by mass spectrometry.
- Fragments are analyzed and graphed to show changes between samples.