產品介紹表觀遺傳學研究組蛋白修飾分析試劑AlphaLISA Epigenetics Reagents

AlphaLISA Epigenetics Reagents

AlphaLISA Epigenetics Reagents 是由 Alpha 技術所延伸出來的便利產品,可適用於偵測 Peptide 或 Full-length 型式的 Histone 蛋白。具有高度專一性,可藉由偵測各種轉譯後修飾類型的 Histone 蛋白,精確定量組蛋白密碼 (histone code) 調控者(例如 Acetyltransferase、Methyltransferase、Deacetylase、Demethylase)的酵素活性。與 ELISA 產品相比,AlphaLISA 偵測試劑具有更加寬廣的偵測區間,可減少樣本稀釋比例的測試步驟;並且具有更精簡快速的實驗流程,讓研究者能夠更快取得定量結果!

偵測原理

AlphaLISA 偵測技術是由兩種核心物質所組成,一為 Donor bead,另一為 Acceptor bead。Donor bead 本身帶有光敏感物質「Phthalocyanine」,因此當以 680 nm 雷射光激發 Donor bead 時,此光敏感物質會催化周遭的氧分子形成活化態,並於作用範圍內激發鄰近的 Acceptor bead 產生化學冷光反應,此冷光能量會進一步轉移至 Acceptor bead 本身所帶有的螢光物質,使其最終產生 615 nm 螢光訊號。與其他偵測平台相比(例如 FRET、TR-FRET),AlphaLISA 技術的能量轉移有效距離長達 200 nm,因此不論是胜肽、重組蛋白、或是細胞內原始含有的蛋白質,皆可作為實驗偵測對象。具有專一性佳、靈敏度高、偵測區間廣泛以及流程簡單等多項優質特點。

在進行實驗時,研究者僅需自行準備 (1). 待測酵素與輔因子 (cofactor)、(2). 鍵結有 Biotin 的 Histone peptide substrate 或 Histone 全長蛋白,即可開始實驗。若是使用試劑盒類型產品偵測細胞內的 Histone 蛋白,則不需額外準備前述兩項材料。歡迎與我們聯繫索取適用產品建議與技術支援。


產品效能

絕佳的專一性!使用 AlphaLISA Di-Methyl-Histone H3 Lysine 4 (H3K4me2) Detection Kit (Cat. No. PK-AL716) 檢測 HeLa 細胞樣本,並以不同類型的 peptide 進行競爭抑制。實驗結果顯示 AlphaLISA 試劑盒的確能夠專一性地辨識偵測細胞內的甲基化組蛋白「H3K4me2」,無論是 H3K4ac peptide、H3K4me3 peptide、H3K9me2 peptide、H3K27me2 peptide、unmodified H3 peptide 皆無法干擾反應。H3K4me1 peptide 雖然會對偵測結果造成影響,但與 H3K4me2 peptide 相比,其結合親和力降低了 30 倍,顯示其並影響性有限。
※ 點擊這裡可瀏覽詳細說明。

 

充分驗證與優化!所有 Alpha 系列的 Histone modification 偵測試劑皆實際使用特定酵素進行完整測試,具有詳盡的測試條件與實驗流程可供參照。例如上圖即是使用 G9a Histone H3-Lysine N-methyltransferase 所進行的酵素用量和時間反應測試結果,受測產品為 Anti-methyl-Histone H3 Lysine 9 (H3K9me2) AlphaLISA Acceptor Beads (Cat. No. PK-AL117) 和 Alpha Streptavidin Donor beads (Cat. No. PK-6760002)。其他測試項目包含:Cofactor titration (ex, SAM)、Enzyme inhibition、Z'-factor determination.
※ 點擊這裡可瀏覽詳細說明。

訂購資訊

目前 Revvity 公司已針對 Histone H3、Histone H4 以及 p53 蛋白質的 Methyl 與 Acetyl 轉譯後修飾推出多款 AlphaLISA 專一性偵測產品。由於產品新增快速,若您在下方列表中找不到需要的產品,歡迎與我們聯繫索取最新即時的產品資訊與技術支援。

AlphaLISA Epigenetic Cellular Detection Kits

Detected Modification Product Name Cat. No.
H3K4 (unmodified) AlphaLISA Unmodified Histone H3 Lysine 4 (H3K4) Detection Kit
This kit is designed to be used as a control for the AlphaLISA Epigenetic Cellular Detection Kits, through detection of unmodified Histone H3 Lysine 4 (H3K4) in cell lysates using a homogeneous AlphaLISA assay (no wash steps).
PK-AL719
H3K4me2 AlphaLISA Di-Methyl-Histone H3 Lysine 4 (H3K4me2) Detection Kit PK-AL716
H3K9ac AlphaLISA Acetylated Histone H3 Lysine 9 (H3K9ac) Detection Kit PK-AL714
H3K27ac AlphaLISA Acetylated-Histone H3 Lysine 27 (H3K27ac) Detection Kit PK-AL720
H3K27me3 AlphaLISA Tri-Methyl-Histone H3 Lysine 27 (H3K27me3) Detection Kit PK-AL722
H3K27me2-1 AlphaLISA Di/Mono-methylated-Histone H3 Lysine 27 (H3K27me2-1) Detection Kit PK-AL721
H3K36me2 AlphaLISA Dimethylated-Histone H3 Lysine 36 (H3K36me2) Detection Kit PK-AL723
H3K79me2 AlphaLISA Dimethylated-Histone H3 Lysine 79 (H3K79me2) Detection Kit PK-AL748
H3K9me2 AlphaLISA Dimethylated-Histone H3 Lysine 9 (H3K9me2) Detection Kit PK-AL717

AlphaLISA Epigenetic Toolbox Reagents (Acceptor Beads)

Detected Modification Product Name Cat. No.
Histone H3 (C-term) AlphaLISA Anti-Histone H3 (C-term) Acceptor Beads
AlphaLISA Acceptor beads designed to capture Histone H3 in a homogeneous AlphaLISA assay for DOT1L. Use in conjunction with biotinylated anti-H3K79me2 (PK-AL148).
PK-AL147
H3K4 (unmodified) AlphaLISA Anti-unmodified Histone H3 Lysine 4 (H3K4) Acceptor Beads PK-AL119
H3K4me1-2 AlphaLISA Anti-methyl-Histone H3 Lysine 4 (H3K4me1-2) Acceptor Beads PK-AL116
H3K9ac AlphaLISA Anti-acetyl-Histone H3 Lysine 9 (H3K9ac) Acceptor Beads PK-AL114
H3K9me2 AlphaLISA Anti-methyl-Histone H3 Lysine 9 (H3K9me2) Acceptor Beads PK-AL117
H3K27me2-1 AlphaLISA Anti-di/mono-methyl-Histone H3 Lysine 27 (H3K27me2-1) Acceptor Beads PK-AL121
H3K27me3 AlphaLISA Anti-tri-methyl-Histone H3 Lysine 27 (H3K27me3) Acceptor Beads PK-AL122
H3K36me2 AlphaLISA Anti-di-methyl-Histone H3 Lysine 36 (H3K36me2) Acceptor Beads (for peptide substrate only) PK-AL123
H3R2me AlphaLISA Anti-methyl Histone H3 Arginine 2 (H3R2me) Acceptor Beads PK-AL139
H3/H4Rme pan AlphaLISA Anti-methyl-Histone H3/H4 Arginine (H3/H4Rme pan) Acceptor Beads PK-AL151
H4 acetyl-Lysine (generic) AlphaLISA Anti-acetyl-Histone H4 Lysine (H4Kac pan) Acceptor Beads PK-AL143
H4R3Me AlphaLISA Anti-methyl-Histone H4 Arginine 3 Acceptor Beads PK-AL150
O-linked GlcNAc AlphaLISA Anti-O-linked GlcNAc Acceptor Beads PK-AL144
p53K382ac AlphaLISA Anti-acetyl-p53 Lysine 382 (p53K382ac) Acceptor Beads PK-AL124

Donor Beads

Product Name Cat. No.
AlphaScreen Streptavidin Donor Beads PK-6760002

Biotinylated Anti-Histone Antibodies

Product Name Cat. No.
AlphaLISA Biotinylated Anti-Histone H3 (C-ter) Antibody
Biotinylated antibody against the C-terminal end of human histone H3, for use in Alpha no-wash epigenetic assays using full-length histone H3 as substrate.
PK-AL118
AlphaLISA Biotin-anti-H3K79me2 antibody
Biotinylated antibody against human histone H3 that is dimethylated on Lysine 79. This antibody is intended for use in AlphaLISA no-wash epigenetic assays using DOT1L methyltransferase with oligonucleosome substrates.
PK-AL148
AlphaLISA Biotin-anti-Histone H4 Antibody
AlphaLISA biotinylated monoclonal antibody recognizing the amino-terminal end of human histone H4. Broad species cross-reactivity is expected based on sequence similarity. This product is designed to be used with AlphaLISA Epigenetics Acceptor beads to detect modified full-length histone H4 in homogeneous AlphaLISA assays.
PK-AL146

Biotinylated Histone H3 or p53 Peptide Substrates

AnaSpec Peptide Cat. No.
Biotinylated Histone H3 peptide (residues 1-21), non-modified (bare)
Sequence: ARTKQTARKSTGGKAPRKQLA-GGK(biotin)-Nh3
61702
Biotinylated Histone H3 peptide (residues 21-44), non-modified (bare)
Sequence: ATKAARKSAPATGGVKKPHRYRPG-GK(biotin)
64440
Biotinylated Histone H3 peptide, acetylated on Lysine 9
Sequence: ARTKQTAR-K(Ac)-STGGKAPRKQLA-GGK(biotin)-Nh3
64361
Biotinylated Histone H3 peptide, acetylated on Lysine 14
Sequence: ARTKQTARKSTGG-K(Ac)-APRKQLA-GGK(biotin)-Nh3
64362
Biotinylated Histone H3 peptide, acetylated on Lysine 9 and Lysine 14
Sequence: ARTKQTAR-K(Ac)-STGG-K(Ac)-APRKQLA-GGK-biotin-Nh3
64363
Biotinylated Histone H3 peptide, acetylated on Lysine 27
Sequence: ATKAAR-K(Ac)-SAPATGGVKKPHRYRPGG-K(biotin)
64846
Biotinylated Histone H3 peptide, mono-methylated on Lysine 4
Sequence: ART-K(Me1)-QTARKSTGGKAPRKQLA-GGK-biotin-Nh3
64355
Biotinylated Histone H3 peptide, di-methylated on Lysine 4
Sequence: ART-K(Me2)-QTARKSTGGKAPRKQLA-GGK(biotin)-Nh3
64356
Biotinylated Histone H3 peptide, tri-methylated on Lysine 4
Sequence: ART-K(Me3)-QTARKSTGGKAPRKQLA-GGK(biotin)-Nh3
64357
Biotinylated Histone H3 peptide, mono-methylated on Lysine 9
Sequence: ARTKQTAR-K(Me1)-STGGKAPRKQLA-GGK(biotin)-Nh3
64358
Biotinylated Histone H3 peptide, di-methylated on Lysine 9
Sequence: ARTKQTAR-K(Me2)-STGGKAPRKQLA-GGK(biotin)-Nh3
64359
Biotinylated Histone H3 peptide, tri-methylated on Lysine 9
Sequence: ARTKQTAR-K(Me3)-STGGKAPRKQLA-GGK(biotin)-Nh3
64360
Biotinylated Histone H3 peptide, mono-methylated on Lysine 27
Sequence: ATKAAR-K(Me1)-SAPATGGVKKPHRYRPG-GK(biotin)
64365
Biotinylated Histone H3 peptide, di-methylated on Lysine 27
Sequence: ATKAAR-K(Me2)-SAPATGGVKKPHRYRPG-GK(biotin)
64366
Biotinylated Histone H3 peptide, tri-methylated on Lysine 27
Sequence: ATKAAR-K(Me3)-SAPATGGVKKPHRYRPG-GK(biotin)
64367
Biotinylated Histone H3 peptide, di-methylated on Lysine 36
Sequence: ATKAARKSAPATGGV-K(Me2)-KPHRYRPG-GK(biotin)
64442
Biotinylated Histone H3 peptide, tri-methylated on Lysine 36
Sequence: ATKAARKSAPATGGV-K(Me3)-KPHRYRPG-GK(biotin)
64441
Biotinylated p53 peptide (residues 361-393), non-modified (bare)
Sequence: Biotin-LC-GSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD-Nh3
62530
Biotinylated p53 peptide, acetylated on Lysine 382
Sequence: Biotin-KGGHLKSKKGQSTSRHK-K(Ac)-LMFKTEGPDSD-Nh3
64869

Other (Buffers)

Product Cat. No.
AlphaLISA Epigenetics Buffer 1, 5X PK-AL008
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