Alpha 技術

Alpha 技術原理

Alpha 技術主要是由兩種核心物質所組成，一為 Donor Bead，另一為 Acceptor Bead。Donor Bead 帶有光敏感物質 "phthalocyanine"，當以 680 nm 雷射光激發 Donor Bead 時，此光敏感物質可催化周遭的氧分子形成活化態，並於 200 nm 作用範圍內，進一步激發鄰近的 Acceptor Bead 產生化學冷光反應，此能量會轉移至 Acceptor Bead 本身所帶有的螢光物質，使其產生出螢光訊號。因此在分析反應中若 Donor Bead 所帶有的 A 分子與 Acceptor Bead 所帶有的 B 分子產生結合或具有交互作用時，Acceptor Bead 即會被帶至 Donor Bead 附近，於 680 nm 雷射光激發下即可測得 Acceptor Bead 所發出的螢光訊號。

Alpha 技術具有兩種 Acceptor Bead，分別為：

1. AlphaScreen Acceptor Bead：螢光訊號波長為 520-620 nm；
2. 新一代的 AlphaLISA Acceptor Bead：螢光訊號波長為 615 nm，相較於舊型的 AlphaScreen Acceptor Bead 具有訊號更強、以及訊號波長區間更集中的優點。

AlphaLISA Acceptor Bead emissions are narrower and brighter than AlphaScreen Acceptor Bead emissions. This allows the development of assays in complex samples such as serum and plasma. (AlphaLISA’s final emission is from europium (615 nm), and AlphaScreen’s final emission is from rubrene (520-620 nm).

Alpha 產品特點

• 偵測靈敏度更高 — Alpha 技術中所使用的 Donor Bead 在 680 nm 雷射光激發下，每秒可催化產生約 60,000 個活化態氧分子，有效達到訊號放大效果，使得系統偵測靈敏度可達 attomole 等級。

• 訊號背景值更低 — Alpha 技術中所使用的 Acceptor Bead，其所激發出的螢光訊號具有長達 0.3 秒的半衰期，因此可以時差性螢光 (Time-Resolved Fluorescence, TRF) 模式偵測訊號，有效降低偵測訊號中的背景值。

• 偵測範圍更廣 — 由於 Donor Bead 與 Acceptor Bead 之間有效的作用距離長達 200 nm，因此無論是小分子 (如：cAMP、cGMP) 或是大分子 (如：Cytokine、Hormone、Kinase、Protein、Whole Cell…) 皆可以 Alpha 技術偵測，且其有效分析範圍 (dynamic range) 更可達 2.5 ~ 5 logs，明顯優於其他傳統分析系統 (如：ELISA, Western Blot)。

• 節省樣本與實驗成本 — 可依據樣本取得的難易度，彈性選擇 96-, 384-, 1536-well format 進行實驗，不但可減少實驗試劑成本，更可避免珍貴樣本的浪費。

• 可直接適用於複雜性樣本 — Alpha 技術可不經額外純化直接適用於複雜性樣本 (如：Cell Lysates、Cell Culture Medium、Serum、Plasma…)，使實驗數據更能真實反應分子在活體內的狀態。

• 隨取隨用，操作更簡便 — Alpha Beads 由於只有 250 nm 直徑大小，因此不像其他 Bead-based 分析平台 (如： Scintillation Proximity Assay, SPA Beads 2-10 µm; FMAT Beads 6-20 µm ) 有珠子容易沈澱的問題，不但可以隨取隨用 (Ready-To-Use, RTU)，而且在使用自動化液體處理系統 (automatic liquid handling device) 分裝試劑時，也不會有阻塞分裝管線的疑慮。

• 符合大量樣本分析時的自動化操作流程規範 — Alpha 技術實驗流程完全不需分離或洗滌步驟，全均質化 (homogeneous) 的實驗流程與簡單的操作步驟 (混和樣本與試劑 → 靜置 → 讀取訊號)，使其非常適合應用於大量樣本的自動化分析 (high throughput screening)。

Alpha Toolbox & AlphaScreen Fusion Detection Kit

Alpha Toolbox 與 AlphaScreen Fusion Detection Kit 系列產品，可以讓研究者自由組合出多種偵測平台，以彈性符合多元化的實驗需求與生物樣本特性。應用實驗包含 Protein:Protein Interaction、DNA:Protein Interaction、DNA Methylation Assay，以及 Receptor-Ligand Binding Assay、Protease Assay、Phosphodiesterase Assay、Biomolecule Detection 等…。歡迎洽詢索取詳細技術資料。

特殊應用範例：

Simultaneous Measurement of Protein Interactions and Phosphorylation — MAP kinase ERK2 dissociation from MAP2K MEK1 upon phosphorylation Assay Type AlphaScreen and AlphaLISA in a single well with recombinant proteins Alpha Donor Beads (GSH-conjugated) AlphaScreen Acceptor Beads (Anti-mouse IgG) AlphaLISA Acceptor Beads (nickel chelate-conjugated) EnVision Multilabel Plate Reader   Results This assay fuses the versatility of AlphaScreen and AlphaLISA platforms. In its unphosphorylated state, GST-ERK2 binds His-MEK1. In the presence of ATP, MEK1 phosphorylates ERK2, causing both proteins to dissociate. Phosphorylation-interaction patterns generated are characteristic for each enzymesubstrate pair and were used to compare mechanisms of action and selectivity of phosphatases as well as small-molecule kinase inhibitors, allowing to discriminate between an ATP-competitor and an allosteric modulator.

訂購資訊 (請點擊下列分類選項，瀏覽詳細訂購資訊)

Alpha Donor Bead

Product Pack Size Cat. No. Anti-FLAG Alpha Donor Beads 1 mg PK-AS103D 5 mg PK-AS103M 25 mg PK-AS103R Anti-Mouse IgG Alpha Donor Beads 1 mg PK-AS104D 5 mg PK-AS104M 25 mg PK-AS104R Anti-Rabbit IgG Alpha Donor Beads 1 mg PK-AS105D 5 mg PK-AS105M 25 mg PK-AS105R AlphaScreen Glutathione Donor Beads 1 mg PK-6765300 5 mg PK-6765301 25 mg PK-6765302 AlphaScreen Nickel Chelate Donor Beads 1 mg PK-AS101D 5 mg PK-AS101M 25 mg PK-AS101R Protein A Alpha Donor Beads 1 mg PK-AS102D 5 mg PK-AS102M 25 mg PK-AS102R Strep-Tactin Alpha Donor Beads 1 mg PK-AS106D 5 mg PK-AS106M 25 mg PK-AS106R AlphaScreen Streptavidin Donor Beads 1 mg PK-6760002S 5 mg PK-6760002 50 mg PK-6760002B AlphaScreen Unconjugated Donor Beads 1 mg PK-6762013 5 mg PK-6762011 50 mg PK-6762012

AlphaLISA Acceptor Bead

Product Pack Size Cat. No. Anti-6xHis AlphaLISA Acceptor Beads 250 µg PK-AL128C 5 mg PK-AL128M 50 mg PK-AL128R Anti-Chicken IgY AlphaLISA Acceptor Beads 250 µg PK-AL131C 5 mg PK-AL131M 50 mg PK-AL131R Anti-c-Myc AlphaLISA Acceptor Beads 250 µg PK-AL111C 5 mg PK-AL111M 50 mg PK-AL111R Anti-DIG AlphaLISA Acceptor Beads 250 µg PK-AL113C 5 mg PK-AL113M 50 mg PK-AL113R Anti-FITC AlphaLISA Acceptor Beads 250 µg PK-AL127C 5 mg PK-AL127M 50 mg PK-AL127R Anti-FLAG AlphaLISA Acceptor Beads 250 µg PK-AL112C 5 mg PK-AL112M 50 mg PK-AL112R Anti-GFP AlphaLISA Acceptor Beads 250 µg PK-AL133C 5 mg PK-AL133M 50 mg PK-AL133R Anti-Goat IgG (Fc specific) AlphaLISA Acceptor Beads 250 µg PK-AL107C 5 mg PK-AL107M 50 mg PK-AL107R Anti-GST AlphaLISA Acceptor Beads 250 µg PK-AL110C 5 mg PK-AL110M 50 mg PK-AL110R Anti-Human IgG (Fc-specific) AlphaLISA Acceptor Beads 250 µg PK-AL103C 5 mg PK-AL103M 50 mg PK-AL103R Anti-Maltose-Binding Protein (MBP) AlphaLISA Acceptor Beads 250 µg PK-AL134C 5 mg PK-AL134M 50 mg PK-AL134R Anti-Mouse IgG (Fc-specific) AlphaLISA Acceptor Beads 250 µg PK-AL105C 5 mg PK-AL105M 50 mg PK-AL105R Anti-Mouse IgM AlphaLISA Acceptor Beads 250 µg PK-AL130C 5 mg PK-AL130M 50 mg PK-AL111R Anti-Rabbit IgG (Fc-specific) AlphaLISA Acceptor Beads 250 µg PK-AL104C 5 mg PK-AL104M 50 mg PK-AL104R Anti-Rat IgG (Fc-specific) AlphaLISA Acceptor Beads 250 µg PK-AL106C 5 mg PK-AL106M 50 mg PK-AL106R Anti-Sheep IgG (Fc specific) AlphaLISA Acceptor Beads 250 µg PK-AL132C 5 mg PK-AL132M 50 mg PK-AL132R Anti-V5 AlphaLISA Acceptor Beads 250 µg PK-AL129C 5 mg PK-AL129M 50 mg PK-AL129R Glutathione AlphaLISA Acceptor Beads 250 µg PK-AL109C 5 mg PK-AL109M 50 mg PK-AL109R Nickel Chelate AlphaLISA Acceptor Beads 250 µg PK-AL108C 5 mg PK-AL108M 50 mg PK-AL108R Protein A AlphaLISA Acceptor Beads 250 µg PK-AL101C 5 mg PK-AL101M 50 mg PK-AL101R Protein G AlphaLISA Acceptor Beads 250 µg PK-AL102C 5 mg PK-AL102M 50 mg PK-AL102R Protein L AlphaLISA Acceptor Beads 250 µg PK-AL126C 5 mg PK-AL126M 50 mg PK-AL126R Strep-Tactin AlphaLISA Acceptor Beads 250 µg PK-AL136C 5 mg PK-AL136M 50 mg PK-AL136R Streptavidin AlphaLISA Acceptor Beads 250 µg PK-AL125C 5 mg PK-AL125M 50 mg PK-AL125R Unconjugated AlphaLISA Acceptor Beads 1 mg PK-6772001 5 mg PK-6772002 50 mg PK-6772003

AlphaScreen Acceptor Bead

Product Pack Size Cat. No. AlphaScreen Protein A Acceptor Beads 5 mg PK-6760137M 50 mg PK-6760137R AlphaScreen Unconjugated Acceptor Beads 1 mg PK-6762003 5 mg PK-6762001 50 mg PK-6762002

AlphaScreen Fusion Tag Detection Kit

Product Pack Size Cat. No. AlphaScreen c-Myc Detection Kit 500 assay PK-6760611C 10,00 assay PK-6760611M 50,00 assay PK-6760611R AlphaScreen DIG (Digoxin/Digoxigenin) Detection Kit 500 assay PK-6760604C 10,00 assay PK-6760604M 50,00 assay PK-6760604R AlphaScreen FITC (Fluorescein) Detection Kit 500 assay PK-6760605C 10,00 assay PK-6760605M 50,00 assay PK-6760605R AlphaScreen FLAG Detection Kit 500 assay PK-6760613C 10,00 assay PK-6760613M 50,00 assay PK-6760613R AlphaScreen GST Detection Kit 500 assay PK-6760603C 10,00 assay PK-6760603M 50,00 assay PK-6760603R AlphaScreen HA (Hemagglutinin) Detection Kit 500 assay PK-6760612C 10,00 assay PK-6760612M 50,00 assay PK-6760612R AlphaScreen Histidine (Nickel Chelate) Detection Kit 500 assay PK-6760619C 10,00 assay PK-6760619M 50,00 assay PK-6760619R AlphaScreen Biotinylated-GST 10,00 assay PK-6760305M AlphaScreen Biotinylated-HIS 10,00 assay PK-6760303M

Other Alpha Products

Classification Product Pack Size Cat. No. AlphaLISA Buffers AlphaLISA Dissociation Buffer, 3X 10 ml PK-AL006C 100 ml PK-AL006F AlphaLISA HiBlock Buffer, 10X 10 ml PK-AL004C 100 ml PK-AL004F AlphaLISA Immunoassay Buffer, 10X 10 ml PK-AL000C 100 ml PK-AL000F AlphaLISA Lysis Buffer, 5X 10 ml PK-AL003C 100 ml PK-AL003F AlphaLISA NaCl Buffer, 5X 10 ml PK-AL007C 100 ml PK-AL007F AlphaLISA Universal Buffer, 5X 10 ml PK-AL001C 100 ml PK-AL001F Bead Conjugation Kit AlphaScreen Conjugation Kit Each Kit Contains: Streptavidin Alpha Donor Bead, unconjugated AlphaScreen Acceptor Bead. 2 x 2 mg PK-6760000K Optimization Tools   AlphaScreen Omnibeads OmniBead are designed as a tool to troubleshoot signal variability. The Bead contain all of the chemical components needed to generate a strong AlphaScreen signal without the presence of AlphaScreen Donor or Acceptor Bead. 1,000 assay PK-6760626D 10,00 assay PK-6760626M 50,00 assay PK-6760626R AlphaScreen TruHits Kit Each Kit Contains: Biotinylated Acceptor Bead, streptavidin Alpha Donor Bead for identifying false positives. 1,000 assay PK-6760627D 10,00 assay PK-6760627M