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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