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AlphaScreen SureFire Cellular Kinase Assay Kits
AlphaScreen® SureFire® 是由 Revvity 公司所研發出的新型態細胞內磷酸化蛋白質偵測技術。此技術幾乎不具有任何技術門檻,只要會使用 Pipetman 就可以進行實驗!流程簡單,從拿到待測檢體開始到取得偵測結果,只要三個步驟就能完成。AlphaScreen SureFire 除了具有無技術門檻以及操作流程簡單的特性之外,還擁有優於 Western Blot、ELISA 的偵測靈敏度與偵測區間,不但可減少樣本用量、提高解析度、並且具有量化的數據顯示方式,彌補了傳統 Western Blot 與 ELISA 的不足之處。
*本系列產品亦可適用於細胞內非磷酸化蛋白質的偵測,例如:GAPDH, Akt, Erk, p53, Topo IIa 等,詳情請見訂購資訊。 |
產品特點:
- 偵測靈敏度高,細胞可不經任何基因轉染 (transfection) 或大量表現 (over-expression) 過程,即可直接進行偵測。
- 適用多種哺乳動物細胞株以及初代細胞 (primary cell)。
- 全均質化 (homogeneous) 操作步驟,流程簡單且不需分離或洗滌步驟,可快速完成偵測實驗。
- 產品種類豐富,涵蓋各種熱門細胞訊息傳遞路徑,如:Akt, JAK/STAT, MAPK, mTOR, NF-kB, TGFb/Smad signaling pathway 等。
實驗步驟 |
Western Blot |
ELISA |
AlphaScreen SureFire |
0 |
取得待測檢體
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取得待測檢體 |
取得待測檢體 |
1 |
電泳 |
加入反應試劑
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加入反應試劑
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2 |
轉漬 |
清洗 x 3次 |
加入偵測試劑 |
3 |
Blocking |
加入偵測抗體 |
偵測 |
4 |
1st 抗體結合 |
清洗 x 3次 |
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5 |
清洗 x 3次 |
加入 2nd 抗體 |
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6 |
2nd 抗體結合 |
清洗 x 3次 |
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7 |
清洗 x 3次 |
加入呈色劑 |
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8 |
呈色反應 |
加入終止反應劑 |
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9 |
偵測 |
偵測 |
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作用原理:
AlphaScreen® SureFire® 作用原理類似 Sandwich ELISA,以下圖「AlphaScreen SureFire p-Erk」偵測試劑為例,當 cell lysates 樣本中含有待測的特定磷酸化蛋白質 "p-Erk" 時,帶有 Erk 專一性抗體的 Donor Bead、與帶有 phospho Erk 專一性抗體的 Acceptor Bead,就能夠經由抗體與標的蛋白間的專一性結合而靠近,此時以 680 nm 雷射光激發 Donor Bead,即可藉由能量轉移反應測得 Acceptor Bead 所發出的 520-620 nm 螢光訊號。
產品效能:
採用 AlphaScreen SureFire 偵測平台可得到量化的實驗結果,讓研究者能夠更清晰地辨識出實驗組 (CTL) 與對照組 (UV) 之間的磷酸化蛋白質表現差異程度。(A) Western blot; (B) AlphaScreen assay for JNK; (C) AlphaScreen SureFire assay for P-p38 and P-Erk2.
[Data source: Apoptosis. 2008 Mar/ 13(3):343-53.] |
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AlphaScreen SureFire 可適用在 Primary Cell 的磷酸化蛋白質偵測。以 AlphaScreen SureFire p70S6K Kit (Cat# PK-TGR70S500) 偵測 Primary Fibroblast Cell 中 phosphorylated p70S6K (Thr389) 的含量,左圖顯示 Rapamycin 對於 p70 S6 Kinase 磷酸化過程具有抑制效果。
[Data source: Cellular and Biochemical mTOR Assays for the Phosphorylation of p70 S6K in Oncology Research.] |
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AlphaScreen SureFire 不需 over-expression 就能直接測得細胞內原始的磷酸化蛋白質含量。以未經刺激活化過的 HUVEC 細胞作為研究對象,並以不同濃度的 U0126 抑制劑 (MEK inhibitor, Calbiochem) 或 LY294002 抑制劑 (PI3K inhibitor, Calbiochem) 分別投與 HUVEC 細胞培養皿中作用,接著以 AlphaScreen SureFire phospho-ERK Assay Kit (Cat# PK-TGRES500) 來偵測 HUVEC 細胞內的磷酸化 ERK1/2 含量。左圖為進行三重複實驗後所得到的實驗結果。結果顯示相較於用來作為對照組的非 MAPK pathway 抑制劑 LY2940022 (▲),U0126 抑制劑 (●) 的確可專一性地抑制 MEK 磷酸化 ERK1/2 的反應,且其抑制活性 IC50: 100 nM 與 1998 年 Duncia et al. 研究團隊發表於 Bioorg Med Chem Lett. 期刊的研究結果相似 (IC50: 70 nM for MEK 1; IC50 : 60 nM for MEK 2)。
[Data source: A Sensitive and Precise Tool for Measuring Endogenous Kinase Activity in Primary Cells — AlphaScreen SureFire.] |
訂購資訊 (請點擊下列分類選項,瀏覽詳細訂購資訊)
AKT Pathway |
Product |
Pack Size |
Cat. No. |
AlphaScreen SureFire (Akt Signaling pathway) research kit |
500 assay |
PK-TGRSP2S500 |
10,000 assay |
PK-TGRSP2S10K |
50,000 assay |
PK-TGRSP2S50K |
AlphaScreen SureFire Akt 1 (p-Ser473) Assay Kit |
500 assay |
PK-TGRA5S500 |
10,000 assay |
PK-TGRA5S10K |
50,000 assay |
PK-TGRA5S50K |
AlphaScreen SureFire Phospho-AKT 1 (Thr308) Kit |
500 assay |
PK-TGRA2S500 |
10,000 assay |
PK-TGRA2S10K |
50,000 assay |
PK-TGRA2S50K |
AlphaScreen SureFire Total Akt1 research kit |
500 assay |
PK-TGRTA1S500 |
10,000 assay |
PK-TGRTA1S10K |
50,000 assay |
PK-TGRTA1S50K |
AlphaScreen SureFire Akt 1/2 (p-Ser473) Assay Kit |
500 assay |
PK-TGRAS500 |
10,000 assay |
PK-TGRAS10K |
50,000 assay |
PK-TGRAS50K |
AlphaScreen SureFire Akt 1/2/3 (p-Ser473) Assay Kit |
100 assay |
PK-TGRA4S100 |
500 assay |
PK-TGRA4S500 |
10,000 assay |
PK-TGRA4S10K |
50,000 assay |
PK-TGRA4S50K |
AlphaScreen SureFire Phospho-AKT 1/2/3 (Thr308) Assay Kit |
100 assay |
PK-TGRA3S100 |
500 assay |
PK-TGRA3S500 |
10,000 assay |
PK-TGRA3S10K |
50,000 assay |
PK-TGRA3S50K |
AlphaScreen SureFire HV™ Total Akt 1/2/3 Assay Kit |
100 assay |
PK-TGRTAPSHV100 |
AlphaScreen SureFire Akt 1/2/3 Total Assay Kit |
100 assay |
PK-TGRTAPS100 |
500 assay |
PK-TGRTAPS500 |
10,000 assay |
PK-TGRTAPS10K |
50,000 assay |
PK-TGRTAPS50K |
AlphaScreen SureFire Phospho-BAD (Ser112) Kit |
500 assay |
PK-TGRBS500 |
10,000 assay |
PK-TGRBS10K |
50,000 assay |
PK-TGRBS50K |
AlphaScreen SureFire Phospho-BAD (Ser136) Kit |
500 assay |
PK-TGRB2S500 |
10,000 assay |
PK-TGRB2S10K |
50,000 assay |
PK-TGRB2S50K |
AlphaScreen SureFire Phospho-Caspase 9 (Ser196) Kit |
500 assay |
PK-TGRC9S500 |
10,000 assay |
PK-TGRC9S10K |
50,000 assay |
PK-TGRC9S50K |
AlphaScreen SureFire Phospho-GSK 3a (Ser21) Kit |
500 assay |
PK-TGRGAS500 |
10,000 assay |
PK-TGRGAS10K |
50,000 assay |
PK-TGRGAS50K |
AlphaScreen SureFire Phospho-GSK 3b (Ser9) Kit |
500 assay |
PK-TGRGBS500 |
10,000 assay |
PK-TGRGBS10K |
50,000 assay |
PK-TGRGBS50K |
AlphaScreen SureFire Phospho-PDK 1 (Ser241) Kit |
500 assay |
PK-TGRPS500 |
10,000 assay |
PK-TGRPS10K |
50,000 assay |
PK-TGRPS50K |
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JAK/STAT Pathway |
Product |
Pack Size |
Cat. No. |
AlphaScreen SureFire STAT1 (p-Tyr701) Assay Kit |
500 assay |
PK-TGRS1S500 |
10,000 assay |
PK-TGRS1S10K |
50,000 assay |
PK-TGRS1S50K |
AlphaScreen SureFire HV™ STAT3 (p-Tyr705) Assay Kit |
100 assay |
PK-TGRS3SHV100 |
AlphaScreen SureFire Phospho-STAT 3 (Tyr705) Assay Kit |
100 assay |
PK-TGRS3S100 |
500 assay |
PK-TGRS3S500 |
10,000 assay |
PK-TGRS3S10K |
50,000 assay |
PK-TGRS3S50K |
AlphaScreen SureFire Phospho-STAT 5 (Tyr694/Tyr699) Assay Kit |
100 assay |
PK-TGRS5S100 |
500 assay |
PK-TGRS5S500 |
10,000 assay |
PK-TGRS5S10K |
50,000 assay |
PK-TGRS5S50K |
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MAPK Pathway |
Product |
Pack Size |
Cat. No. |
AlphaScreen SureFire ATF1 (p-Ser63) Assay Kit |
500 assay |
PK-TGRAF1S500 |
10,000 assay |
PK-TGRAF1S10K |
50,000 assay |
PK-TGRAF1S50K |
AlphaScreen SureFire Phospho-ATF2 (Thr71) Assay Kit |
500 assay |
PK-TGRAF2S500 |
10,000 assay |
PK-TGRAF2S10K |
50,000 assay |
PK-TGRAF2S50K |
AlphaScreen SureFire cJun (p-Ser63) Assay Kit |
500 assay |
PK-TGRCJS500 |
10,000 assay |
PK-TGRCJS10K |
50,000 assay |
PK-TGRCJS50K |
AlphaScreen SureFire cJun (p-Ser73) Assay Kit |
500 assay |
PK-TGRCJ2S500 |
10,000 assay |
PK-TGRCJ2S10K |
50,000 assay |
PK-TGRCJ2S50K |
AlphaScreen SureFire CREB (p-Ser133) Assay Kit |
500 assay |
PK-TGRCBS500 |
10,000 assay |
PK-TGRCBS10K |
50,000 assay |
PK-TGRCBS50K |
AlphaScreen SureFire ELK1 (p-Ser383) Assay Kit |
500 assay |
PK-TGRELKS500 |
10,000 assay |
PK-TGRELKS10K |
50,000 assay |
PK-TGRELKS50K |
AlphaScreen SureFire HV™ ERK 1/2 (p-Thr202/Tyr204) Assay Kit |
100 assay |
PK-TGRESHV100 |
AlphaScreen SureFire Phospho-ERK 1/2 Assay Kit |
500 assay |
PK-TGRES500 |
10,000 assay |
PK-TGRES10K |
50,000 assay |
PK-TGRES50K |
AlphaScreen SureFire HV™ ERK 1/2 Total Assay Kit |
100 assay |
PK-TGRTESHV100 |
AlphaScreen SureFire ERK 1/2 Total Assay Kit |
500 assay |
PK-TGRTES500 |
10,000 assay |
PK-TGRTES10K |
50,000 assay |
PK-TGRTES50K |
AlphaScreen SureFire Phospho-HSP27 (Ser82) Assay Kit |
500 assay |
PK-TGRHS27S500 |
10,000 assay |
PK-TGRHS27S10K |
50,000 assay |
PK-TGRHS27S50K |
AlphaScreen SureFire JNK 1/2/3 (p-Thr183/Tyr185) Assay Kit |
500 assay |
PK-TGRJPS500 |
10,000 assay |
PK-TGRJPS10K |
50,000 assay |
PK-TGRJPS50K |
AlphaScreen SureFire JNK 1/2/3 Total Assay Kit |
500 assay |
PK-TGRTJS500 |
10,000 assay |
PK-TGRTJS10K |
50,000 assay |
PK-TGRTJS50K |
AlphaScreen SureFire JNK 1/3 (p-Thr183/Tyr185) Assay Kit |
100 assay |
PK-TGRJS100 |
500 assay |
PK-TGRJS500 |
10,000 assay |
PK-TGRJS10K |
50,000 assay |
PK-TGRJS50K |
AlphaScreen SureFire MAPK pathway research kit |
500 assay |
PK-TGRSP1S500 |
10,000 assay |
PK-TGRSP1S10K |
50,000 assay |
PK-TGRSP1S50K |
AlphaScreen SureFire Phospho-MEK 1 Kit |
500 assay |
PK-TGRMS500 |
10,000 assay |
PK-TGRMS10K |
50,000 assay |
PK-TGRMS50K |
AlphaScreen SureFire MKK3 (p-Ser189), MKK6 (p-Ser207) Assay Kit |
500 assay |
PK-TGRMK3S500 |
10,000 assay |
PK-TGRMK3S10K |
50,000 assay |
PK-TGRMK3S50K |
AlphaScreen SureFire MKK4 (p-Ser257/Thr261) Assay Kit |
500 assay |
PK-TGRMK4S500 |
10,000 assay |
PK-TGRMK4S10K |
50,000 assay |
PK-TGRMK4S50K |
AlphaScreen SureFire Phospho-p38 MAPK Assay Kit |
100 assay |
PK-TGR38S100 |
500 assay |
PK-TGR38S500 |
10,000 assay |
PK-TGR38S10K |
50,000 assay |
PK-TGR38S50K |
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mTOR Pathway |
Product |
Pack Size |
Cat. No. |
AlphaScreen SureFire 4EBP1 (p-Thr37/Thr46) Assay Kit Kit |
500 assay |
PK-TGR4ES500 |
10,000 assay |
PK-TGR4ES10K |
50,000 assay |
PK-TGR4ES50K |
AlphaScreen SureFire 4EBP1 (p-Thr70) Assay Kit |
500 assay |
PK-TGR4E2S500 |
10,000 assay |
PK-TGR4E2S10K |
50,000 assay |
PK-TGR4E2S50K |
AlphaScreen SureFire EIF4E (p-Ser209) Assay Kit |
500 assay |
PK-TGREIF4S500 |
10,000 assay |
PK-TGREIF4S10K |
50,000 assay |
PK-TGREIF4S50K |
AlphaScreen SureFire mTOR (p-Ser2448) |
500 assay |
PK-TGRMTS500 |
10,000 assay |
PK-TGRMTS10K |
50,000 assay |
PK-TGRMTS50K |
AlphaScreen SureFire Phospho-mTOR (Ser2481) Kit |
500 assay |
PK-TGRMT2S500 |
10,000 assay |
PK-TGRMT2S10K |
50,000 assay |
PK-TGRMT2S50K |
AlphaScreen SureFire p70 S6K (p-Thr229) Assay Kit |
500 assay |
PK-TGR703S500 |
10,000 assay |
PK-TGR703S10K |
50,000 assay |
PK-TGR703S50K |
AlphaScreen SureFire phospho-p70 S6K (Thr389) Assay Kit |
100 assay |
PK-TGR70S100 |
500 assay |
PK-TGR70S500 |
10,000 assay |
PK-TGR70S10K |
50,000 assay |
PK-TGR70S50K |
AlphaScreen SureFire p70 S6K (p-Thr421/Ser424) Assay Kit |
500 assay |
PK-TGR702S500 |
10,000 assay |
PK-TGR702S10K |
50,000 assay |
PK-TGR702S50K |
AlphaScreen SureFire S6RP (p-Ser235/Ser236) Assay Kit |
500 assay |
PK-TGRS6PS500 |
10,000 assay |
PK-TGRS6PS10K |
50,000 assay |
PK-TGRS6PS50K |
AlphaScreen SureFire S6RP (p-Ser240/Ser244) Assay Kit |
500 assay |
PK-TGRS6P2S500 |
10,000 assay |
PK-TGRS6P2S10K |
50,000 assay |
PK-TGRS6P2S50K |
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NFkB Pathway |
Product |
Pack Size |
Cat. No. |
AlphaScreen SureFire IkBalpha (p-Ser32/Ser36) Assay Kit |
500 assay |
PK-TGRIKS500 |
10,000 assay |
PK-TGRIKS10K |
50,000 assay |
PK-TGRIKS50K |
AlphaScreen SureFire Total IkBalpha Assay Kit |
500 assay |
PK-TGRTIKS500 |
10,000 assay |
PK-TGRTIKS10K |
50,000 assay |
PK-TGRTIKS50K |
AlphaScreen SureFire IKKa (p-Ser176/Ser180) Assay Kit |
500 assay |
PK-TGRKAS500 |
10,000 assay |
PK-TGRKAS10K |
50,000 assay |
PK-TGRKAS50K |
AlphaScreen SureFire IKKb (p-Ser177/Ser181) Assay Kit |
500 assay |
PK-TGRKBS500 |
10,000 assay |
PK-TGRKBS10K |
50,000 assay |
PK-TGRKBS50K |
AlphaScreen SureFire NFkB Signaling pathway research kit |
500 assay |
PK-TGRSP3S500 |
10,000 assay |
PK-TGRSP3S10K |
50,000 assay |
PK-TGRSP3S50K |
AlphaScreen SureFire HV™ NFkΒ p65 (p-Ser536) Assay Kit |
100 assay |
PK-TGRNFSHV100 |
AlphaScreen SureFire NFkB (p-Ser536) Assay Kit |
500 assay |
PK-TGRNFS500 |
10,000 assay |
PK-TGRNFS10K |
50,000 assay |
PK-TGRNFS50K |
AlphaScreen SureFire Total NFkB p65 Assay Kit |
500 assay |
PK-TGRTNFS500 |
10,000 assay |
PK-TGRTNFS10K |
50,000 assay |
PK-TGRTNFS50K |
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Others |
Product |
Pack Size |
Cat. No. |
AlphaScreen SureFire ALK (p-Tyr1586) Assay Kit |
500 assay |
PK-TGRALS500 |
10,000 assay |
PK-TGRALS10K |
50,000 assay |
PK-TGRALS50K |
AlphaScreen SureFire ALK (p-Tyr1604) Assay Kit |
500 assay |
PK-TGRAL2S500 |
10,000 assay |
PK-TGRAL2S10K |
50,000 assay |
PK-TGRAL2S50K |
AlphaScreen SureFire ALK Total Assay Kit |
500 assay |
PK-TGRTALS500 |
10,000 assay |
PK-TGRTALS10K |
50,000 assay |
PK-TGRTALS50K |
AlphaScreen SureFire Phospho-beta Catenin (Ser45) Assay Kit |
10,000 assay |
PK-TGRCHK1S10K |
50,000 assay |
PK-TGRCHK1S50K |
AlphaScreen SureFire CHK1 (p-Ser345) Assay Kit |
500 assay |
PK-TGRCHK1S500 |
10,000 assay |
PK-TGRCHK1S10K |
50,000 assay |
PK-TGRCHK1S50K |
AlphaScreen SureFire CHK2 (p-Thr68) Assay Kit |
500 assay |
PK-TGRCHK2S500 |
10,000 assay |
PK-TGRCHK2S10K |
50,000 assay |
PK-TGRCHK2S50K |
AlphaScreen SureFire Phospho-Cofilin (Ser3) Assay Kit |
500 assay |
PK-TGRCOFS500 |
10,000 assay |
PK-TGRCOFS10K |
50,000 assay |
PK-TGRCOFS50K |
AlphaScreen SureFire EGFR (p-Tyr1068) Assay Kit |
500 assay |
PK-TGRERS500 |
10,000 assay |
PK-TGRERS10K |
50,000 assay |
PK-TGRERS50K |
AlphaScreen SureFire Phospho-eIF2a (Ser51) Assay Kit |
500 assay |
PK-TGREIF2S500 |
10,000 assay |
PK-TGREIF2S10K |
50,000 assay |
PK-TGREIF2S50K |
AlphaScreen SureFire ErbB2 (p-Tyr1221/Tyr1222) Assay Kit |
500 assay |
PK-TGREB2S500 |
10,000 assay |
PK-TGREB2S10K |
50,000 assay |
PK-TGREB2S50K |
AlphaScreen SureFire GAPDH Assay Kit |
100 assay |
PK-TGRGDS100 |
500 assay |
PK-TGRGDS500 |
10,000 assay |
PK-TGRGDS10K |
50,000 assay |
PK-TGRGDS50K |
AlphaScreen SureFire Histone H3 (p-Ser10) Assay Kit |
500 assay |
PK-TGRH3S500 |
10,000 assay |
PK-TGRH3S10K |
50,000 assay |
PK-TGRH3S50K |
AlphaScreen SureFire IGFR (p-Tyr1135/1136) Assay Kit |
500 assay |
PK-TGRIGFS500 |
10,000 assay |
PK-TGRIGFS10K |
50,000 assay |
PK-TGRIGFS50K |
AlphaScreen SureFire InsulinR (p-Tyr1150/Tyr1151) Assay Kit |
500 assay |
PK-TGRIRS500 |
10,000 assay |
PK-TGRIRS10K |
50,000 assay |
PK-TGRIRS50K |
AlphaScreen SureFire HV™ p53 (p-Ser15) Assay Kit |
100 assay |
PK-TGRP53SHV100 |
AlphaScreen SureFire p53 (p-Ser15) Assay Kit |
500 assay |
PK-TGRP53S500 |
10,000 assay |
PK-TGRP53S10K |
50,000 assay |
PK-TGRP53S50K |
AlphaScreen SureFire Phospho-p53 (Ser392) Assay Kit |
500 assay |
PK-TGRP532S500 |
10,000 assay |
PK-TGRP532S10K |
50,000 assay |
PK-TGRP532S50K |
AlphaScreen SureFire HV™ Total p53 Assay Kit |
100 assay |
PK-TGRT53SHV100 |
AlphaScreen SureFire p53 Total Assay Kit |
500 assay |
PK-TGRT53S500 |
10,000 assay |
PK-TGRT53S10K |
50,000 assay |
PK-TGRT53S50K |
AlphaScreenSureFire SMAD1 (p-Ser463/Ser465) Assay Kit |
500 assay |
PK-TGRSM1S500 |
10,000 assay |
PK-TGRSM1S10K |
50,000 assay |
PK-TGRSM1S50K |
AlphaScreenSureFire SMAD2 (p-Ser465/Ser467) Assay Kit |
500 assay |
PK-TGRSM2S500 |
10,000 assay |
PK-TGRSM2S10K |
50,000 assay |
PK-TGRSM2S50K |
AlphaScreenSureFire p-Smad3 (Ser423/425) research kit |
500 assay |
PK-TGRSM3S500 |
10,000 assay |
PK-TGRSM3S10K |
50,000 assay |
PK-TGRSM3S50K |
AlphaScreen SureFire Phospho-Topo IIa (Ser1106) Assay Kit |
500 assay |
PK-TGRT2A2S500 |
10,000 assay |
PK-TGRT2A2S10K |
50,000 assay |
PK-TGRT2A2S50K |
AlphaScreen SureFire Phospho-Topo IIa (Thr1343) Assay Kit |
500 assay |
PK-TGRT2AS500 |
10,000 assay |
PK-TGRT2AS10K |
50,000 assay |
PK-TGRT2AS50K |
AlphaScreen SureFire Total Topo IIa Assay Kit |
500 assay |
PK-TGRTT2AS500 |
10,000 assay |
PK-TGRTT2AS10K |
50,000 assay |
PK-TGRTT2AS50K |
AlphaScreen SureFire Phospho-VEGF2 Receptor (Tyr1175) research kit |
500 assay |
PK-TGRVRS500 |
10,000 assay |
PK-TGRVRS10K |
50,000 assay |
PK-TGRVRS50K |
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