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ADP-Glo™ キナーゼアッセイサポート
キナーゼ反応から産生される ADP を測定する発光ベースのADP-Glo™ Kinase Assay（Promega社）はシンプルなプロトコールで様々なキナーゼ活性を検出することが出来る高感度ルミネッセンスアッセイで、キナーゼをターゲットとした研究や薬剤開発において非常に利便性の高いプラットフォームです。
カルナでは、お客様のラボでADP-Glo™ Kinase Assayを用いた薬剤評価やスクリーニングの為のアッセイを立ち上げる際のご参考情報として、自社製タンパク質製品を用いて測定したADP-Glo™アッセイの実施条件を公開しております。
このページで紹介されているアッセイ条件で取得されたデータはActivity Data欄添付のPDFにてご覧頂けます。ターゲット毎のATP濃度、金属イオン条件、添加物条件、反応時間などの詳細は各添付PDF内でご確認ください。
これらデータ取得に用いられたキナーゼタンパク質、ビオチン標識ペプチド基質、アッセイバッファー製品は全て当社で提供中です。製品のお問い合わせ、ご質問、ご相談等はこちらよりまたはinfo@carnabio.com からご連絡ください。
ADP-Glo™ Kinase Assay 反応条件一覧表
| Kinase Name | Protein Kinase | Substrate | Assay Buffer |
ATPase Activity |
Sample Activity Data |
||||
|---|---|---|---|---|---|---|---|---|---|
| Tag | Carna Product Name | Cat. No. | Carna Substrate Name | Cat. No. | Conc. (uM) |
Cat. No. | H:High M:Moderate L:Low |
||
| ABL(ABL1) | HIS | ABL(ABL1) | 08-001 | Biotin-ABLtide | BTSU16 | 10 | ASBF01 | L | |
| ABL(ABL1)[E255K] | HIS | ABL(ABL1)[E255K] | 08-094 | Biotin-ABLtide | BTSU16 | 10 | ASBF01 | L | |
| ABL(ABL1)[T315I] | HIS | ABL(ABL1)[T315I] | 08-093 | Biotin-ABLtide | BTSU16 | 10 | ASBF01 | L | |
| ACK(TNK2) | GST | ACK(TNK2) | 08-196 | Biotin-WASP peptide | BTSU39 | 50 | ASBF01 | L | |
| AKT1 | GST | AKT1 | 01-101 | Biotin-Crosstide | BTSU19 | 10 | ASBF01 | L | |
| AKT2 | GST | AKT2 | 01-102 | Biotin-Crosstide | BTSU19 | 10 | ASBF01 | L | |
| AKT3 | GST | AKT3 | 01-103 | Biotin-Crosstide | BTSU19 | 10 | ASBF01 | L | |
| ALK | GST | ALK | 08-518 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| ALK[C1156Y] | GST | ALK[C1156Y] | 08-530 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| ALK[F1174L] | GST | ALK[F1174L] | 08-519 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| ALK[G1202R] | GST | ALK[G1202R] | 08-544 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| ALK[G1269A] | GST | ALK[G1269A] | 08-537 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| ALK[L1196M] | GST | ALK[L1196M] | 08-529 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| ALK[R1275Q] | GST | ALK[R1275Q] | 08-520 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| ALK[T1151_L1152insT] | GST | ALK[T1151_L1152insT] | 08-539 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| AMPKα1/β1/γ1 | GST | AMPKα1/β1/γ1 | 02-113 | Biotin-SAMS peptide | BTSU26 | 10 | ASBF01 | L | |
| AMPKα2/β1/γ1 | GST | AMPKα2/β1/γ1 | 02-114 | Biotin-SAMS peptide | BTSU26 | 10 | ASBF01 | L | |
| ARG(ABL2) | GST | ARG(ABL2) | 08-102 | Biotin-ABLtide | BTSU16 | 10 | ASBF01 | L | |
| AurA | GST | AurA | 05-101 | Biotin-Kemptide | BTSU09 | 10 | ASBF01 | L | |
| AurB(AURKB)／INCENP | GST | AurB(AURKB)／INCENP | 05-102 | Biotin-Kemptide | BTSU09 | 10 | ASBF01 | M | |
| AurC | GST | AurC | 05-103 | Biotin-Kemptide | BTSU09 | 10 | ASBF01 | L | |
| AXL | GST | AXL | 08-107 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| BLK | GST | BLK | 08-164 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| BMX | GST | BMX | 08-179 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| BRK(PTK6) | GST | BRK(PTK6) | 08-165 | Biotin-Blk/Lyntide | BTSU04 | 50 | ASBF01 | L | |
| BRSK1 | GST | BRSK1 | 02-115 | Biotin-CHKtide | BTSU08 | 10 | ASBF01 | L | |
| BRSK2 | GST | BRSK2 | 02-116 | Biotin-CHKtide | BTSU08 | 10 | ASBF01 | L | |
| BTK | GST | BTK | 08-180 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| BTK[C481S] | GST | BTK[C481S] | 08-547 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| BUB1／BUB3 | GST | BUB1／BUB3 | 05-187 | Biotin-H2A peptide | BTSU47 | 10 | ASBF01 | L | |
| CaMK1α | GST | CaMK1α | 02-104 | Biotin-GS peptide | BTSU05 | 10 | ASBF01 | L | |
| CaMK1δ | GST | CaMK1δ | 02-106 | Biotin-Synapsin peptide | BTSU45 | 10 | ASBF01 | L | |
| CaMK2α(CAMK2A) | GST | CaMK2α(CAMK2A) | 02-109 | Biotin-GS peptide | BTSU05 | 10 | ASBF01 | L | |
| CaMK2β(CAMK2B) | GST | CaMK2β(CAMK2B) | 02-110 | Biotin-GS peptide | BTSU05 | 10 | ASBF01 | L | |
| CaMK2γ(CAMK2G) | GST | CaMK2γ(CAMK2G) | 02-112 | Biotin-GS peptide | BTSU05 | 10 | ASBF01 | L | |
| CaMK2δ(CAMK2D) | GST | CaMK2δ(CAMK2D) | 02-111 | Biotin-GS peptide | BTSU05 | 10 | ASBF01 | L | |
| CaMK4 | GST | CaMK4 | 02-108 | Biotin-GS peptide | BTSU05 | 10 | ASBF01 | L | |
| CDC7／ASK | GST | CDC7／ASK | 05-109 | Biotin-MCM2 peptide | BTSU33 | 10 | ASBF01 | H | |
| CDK1(CDC2)／CycB1 | GST | CDK1(CDC2)／CycB1 | 04-102 | Biotin-Modified Histone H1 | BTSU13 | 10 | ASBF01 | L | |
| CDK2／CycA2 | GST | CDK2／CycA2 | 04-103 | Biotin-Modified Histone H1 | BTSU13 | 10 | ASBF01 | L | |
| CDK2／CycE1 | GST | CDK2／CycE1 | 04-165 | Biotin-Modified Histone H1 | BTSU13 | 10 | ASBF01 | L | |
| CDK3／CycE1 | GST | CDK3／CycE1 | 04-104 | Biotin-Modified Histone H1 | BTSU13 | 10 | ASBF01 | L | |
| CDK4／CycD1 | GST | CDK4／CycD1 | 04-120 | Biotin-DYRKtide | BTSU06 | 10 | ASBF01 | L | |
| CDK4／CycD3 | GST | CDK4／CycD3 | 04-105 | Biotin-DYRKtide | BTSU06 | 10 | ASBF01 | L | |
| CDK5／p25 | GST | CDK5／p25 | 04-106 | Biotin-Modified Histone H1 | BTSU13 | 10 | ASBF01 | L | |
| CDK6／CycD3 | GST | CDK6／CycD3 | 04-107 | Biotin-DYRKtide | BTSU06 | 50 | ASBF01 | L | |
| CDK7／CycH／MAT1 | GST | CDK7／CycH／MAT1 | 04-108 | Biotin-CTD3 peptide | BTSU30 | 50 | ASBF01 | L | |
| CDK9／CycT1 | GST | CDK9／CycT1 | 04-110 | Biotin-CDK9 substrate | BTSU29 | 50 | ASBF01 | L | |
| CGK2 | GST | CGK2 | 01-143 | Biotin-Kemptide | BTSU09 | 10 | ASBF01 | L | |
| CHK1(CHEK1) | GST | CHK1(CHEK1) | 02-117 | Biotin-CHKtide | BTSU08 | 10 | ASBF01 | L | |
| CHK2(CHEK2) | GST | CHK2(CHEK2) | 02-162 | Biotin-CHKtide | BTSU08 | 10 | ASBF01 | L | |
| CK1α | GST | CK1α | 03-101 | Biotin-CKtide | BTSU12 | 50 | ASBF01 | L | |
| CK1γ1 | GST | CK1γ1 | 03-105 | Biotin-CKtide | BTSU12 | 10 | ASBF01 | L | |
| CK1γ2 | GST | CK1γ2 | 03-106 | Biotin-CKtide | BTSU12 | 10 | ASBF01 | L | |
| CK1γ3 | GST | CK1γ3 | 03-107 | Biotin-CKtide | BTSU12 | 10 | ASBF01 | L | |
| CK1δ | GST | CK1δ | 03-103 | Biotin-CKtide | BTSU12 | 10 | ASBF01 | L | |
| CK1ε | GST | CK1ε | 03-104 | Biotin-CKtide | BTSU12 | 50 | ASBF01 | L | |
| CK2α1/β | GST | CK2α1/β | 05-184 | Biotin-CK2tide | BTSU25 | 50 | ASBF01 | L | |
| CK2α2/β | GST | CK2α2/β | 05-185 | Biotin-CK2tide | BTSU25 | 50 | ASBF01 | L | |
| CLK1 | GST | CLK1 | 04-126 | Biotin-DYRKtide | BTSU06 | 50 | ASBF01 | M | |
| CLK2 | GST | CLK2 | 04-127 | Biotin-DYRKtide | BTSU06 | 10 | ASBF01 | L | |
| CLK3 | GST | CLK3 | 04-128 | Biotin-DYRKtide | BTSU06 | 50 | ASBF01 | L | |
| CRIK | GST | CRIK | 01-104 | Biotin-Histone H3 peptide | BTSU23 | 10 | ASBF01 | L | |
| CSK | GST | CSK | 08-111 | Biotin-Srctide | BTSU01 | 50 | ASBF01 | L | |
| DAPK1 | GST | DAPK1 | 02-134 | Biotin-DAPK1tide | BTSU17 | 10 | ASBF01 | M | |
| DCAMKL1 | GST | DCAMKL1 | 02-139 | Biotin-GS peptide | BTSU05 | 50 | ASBF01 | L | |
| DCAMKL2 | GST | DCAMKL2 | 02-140 | Biotin-GS peptide | BTSU05 | 10 | ASBF01 | L | |
| DDR1 | GST | DDR1 | 08-113 | Biotin-IRS1 | BTSU07 | 50 | ASBF01 | L | |
| DDR2 | GST | DDR2 | 08-114 | Biotin-IRS1 | BTSU07 | 10 | ASBF01 | L | |
| DYRK1A | GST | DYRK1A | 04-130 | Biotin-DYRKtide | BTSU06 | 10 | ASBF01 | L | |
| DYRK1B | GST | DYRK1B | 04-131 | Biotin-DYRKtide | BTSU06 | 10 | ASBF01 | L | |
| DYRK2 | GST | DYRK2 | 04-132 | Biotin-DYRKtide | BTSU06 | 10 | ASBF01 | L | |
| DYRK3 | GST | DYRK3 | 04-133 | Biotin-DYRKtide | BTSU06 | 10 | ASBF01 | L | |
| EEF2K | GST | EEF2K | 10-113 | Biotin-EEF2Ktide | BTSU40 | 10 | ASBF01 | L | |
| EGFR | GST | EGFR | 08-115 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| EGFR[C797S／L858R] | GST | EGFR[C797S／L858R] | 08-563 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| EGFR[d746-750] | GST | EGFR[d746-750] | 08-527 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| EGFR[d746-750／C797S] | GST | EGFR[d746-750／C797S] | 08-564 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| EGFR[d746-750／T790M] | GST | EGFR[d746-750／T790M] | 08-528 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| EGFR[d746-750／T790M／C797S] | GST | EGFR[d746-750／T790M／C797S] | 08-565 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| EGFR[D770_N771insNPG] | GST | EGFR[D770_N771insNPG] | 08-553 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| EGFR[L858R] | GST | EGFR[L858R] | 08-502 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| EGFR[L861Q] | GST | EGFR[L861Q] | 08-513 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| EGFR[T790M] | GST | EGFR[T790M] | 08-194 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| EGFR[T790M／C797S／L858R] | GST | EGFR[T790M／C797S／L858R] | 08-559 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | M | |
| EGFR[T790M／L858R] | GST | EGFR[T790M／L858R] | 08-510 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| EML4-ALK | GST | EML4-ALK | 08-516 | Biotin-Srctide | BTSU01 | 50 | ASBF01 | L | |
| EPHA1 | GST | EPHA1 | 08-119 | Biotin-Blk/Lyntide | BTSU04 | 10 | ASBF01 | L | |
| EPHA2 | GST | EPHA2 | 08-121 | Biotin-Blk/Lyntide | BTSU04 | 10 | ASBF01 | L | |
| EPHA3 | GST | EPHA3 | 08-122 | Biotin-Blk/Lyntide | BTSU04 | 10 | ASBF01 | L | |
| EPHA4 | GST | EPHA4 | 08-123 | Biotin-Blk/Lyntide | BTSU04 | 10 | ASBF01 | L | |
| EPHA5 | GST | EPHA5 | 08-124 | Biotin-Blk/Lyntide | BTSU04 | 10 | ASBF01 | L | |
| EPHA6 | GST | EPHA6 | 08-125 | Biotin-Blk/Lyntide | BTSU04 | 10 | ASBF01 | L | |
| EPHA7 | GST | EPHA7 | 08-126 | Biotin-Blk/Lyntide | BTSU04 | 10 | ASBF01 | L | |
| EPHA8 | GST | EPHA8 | 08-127 | Biotin-Blk/Lyntide | BTSU04 | 10 | ASBF01 | L | |
| EPHB1 | GST | EPHB1 | 08-128 | Biotin-Blk/Lyntide | BTSU04 | 10 | ASBF01 | L | |
| EPHB2 | GST | EPHB2 | 08-129 | Biotin-Blk/Lyntide | BTSU04 | 10 | ASBF01 | L | |
| EPHB3 | GST | EPHB3 | 08-130 | Biotin-Blk/Lyntide | BTSU04 | 10 | ASBF01 | L | |
| EPHB4 | GST | EPHB4 | 08-131 | Biotin-Blk/Lyntide | BTSU04 | 10 | ASBF01 | L | |
| Erk1(MAPK3) | GST | Erk1(MAPK3) | 04-142 | Biotin-Modified Erktide | BTSU03 | 10 | ASBF01 | L | |
| Erk2(MAPK1) | GST | Erk2(MAPK1) | 04-143 | Biotin-Modified Erktide | BTSU03 | 10 | ASBF01 | L | |
| Erk5(MAPK7) | GST | Erk5(MAPK7) | 04-146 | Biotin-EGFR-derived peptide | BTSU31 | 10 | ASBF01 | H | |
| FAK(PTK2) | GST | FAK(PTK2) | 08-137 | Biotin-Blk/Lyntide | BTSU04 | 50 | ASBF01 | L | |
| FER | GST | FER | 08-139 | Biotin-Srctide | BTSU01 | 50 | ASBF01 | L | |
| FES | GST | FES | 08-140 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| FGFR1 | GST | FGFR1 | 08-133 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| FGFR1[V561M] | GST | FGFR1[V561M] | 08-536 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| FGFR2 | GST | FGFR2 | 08-134 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| FGFR2[V564I] | GST | FGFR2[V564I] | 08-546 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| FGFR3 | GST | FGFR3 | 08-135 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| FGFR3[K650E] | GST | FGFR3[K650E] | 08-501 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| FGFR3[K650M] | GST | FGFR3[K650M] | 08-199 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| FGFR3[V555L] | GST | FGFR3[V555L] | 08-548 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| FGFR3[V555M] | GST | FGFR3[V555M] | 08-543 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| FGFR4 | GST | FGFR4 | 08-136 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| FGFR4[N535K] | GST | FGFR4[N535K] | 08-524 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| FGFR4[V550E] | GST | FGFR4[V550E] | 08-525 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| FGFR4[V550L] | GST | FGFR4[V550L] | 08-526 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| FGR | GST | FGR | 08-166 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| FLT1 | GST | FLT1 | 08-189 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| FLT3 | GST | FLT3 | 08-154 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| FLT4 | GST | FLT4 | 08-190 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| FMS(CSF1R) | GST | FMS(CSF1R) | 08-155 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| FRK | GST | FRK | 08-167 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| FYN [isoform a] | GST | FYN [isoform a] | 08-168 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| FYN [isoform b] | GST | FYN [isoform b] | 08-531 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| GSK3α | GST | GSK3α | 04-140 | Biotin-CREBtide-p | BTSU22 | 10 | ASBF01 | L | |
| GSK3β(GSK3B) | GST | GSK3β(GSK3B) | 04-141 | Biotin-CREBtide-p | BTSU22 | 10 | ASBF01 | L | |
| Haspin | GST | Haspin | 05-111 | Biotin-Histone H3 peptide | BTSU23 | 10 | ASBF01 | L | |
| HCK | GST | HCK | 08-169 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| HER2(ERBB2) | HIS | HER2(ERBB2) | 08-016 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| HER4 | GST | HER4 | 08-118 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| HGK | GST | HGK | 07-137 | Biotin-Moesin-derived peptide | BTSU18 | 10 | ASBF01 | L | |
| HIPK1 | GST | HIPK1 | 04-135 | Biotin-DYRKtide | BTSU06 | 50 | ASBF01 | L | |
| HIPK2 | GST | HIPK2 | 04-136 | Biotin-DYRKtide | BTSU06 | 50 | ASBF01 | L | |
| HIPK3 | GST | HIPK3 | 04-137 | Biotin-DYRKtide | BTSU06 | 50 | ASBF01 | L | |
| HIPK4 | GST | HIPK4 | 04-138 | Biotin-DYRKtide | BTSU06 | 50 | ASBF01 | L | |
| HPK1(MAP4K1) | DYKDDDDK | HPK1(MAP4K1) | 07-410 | Biotin-S6K2 peptide | BTSU14 | 10 | ASBF01 | L | |
| IGF1R | GST | IGF1R | 08-141 | Biotin-IRS1 | BTSU07 | 10 | ASBF01 | L | |
| IKKβ | HIS | IKKβ | 05-084 | Biotin-Modified IκBα-derived peptide | BTSU34 | 50 | ASBF01 | H | |
| IKKε | GST | IKKε | 05-114 | Biotin-IκBα peptide | BTSU42 | 50 | ASBF01 | H | |
| INSR | GST | INSR | 08-142 | Biotin-IRS1 | BTSU07 | 10 | ASBF01 | L | |
| IRR | GST | IRR | 08-143 | Biotin-IRS1 | BTSU07 | 10 | ASBF01 | L | |
| ITK | GST | ITK | 08-181 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | H | |
| JAK1 | GST | JAK1 | 08-144 | Biotin-JAK1 substrate peptide | BTSU32 | 50 | ASBF01 | L | |
| JAK2 | HIS | JAK2 | 08-045 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| JAK3 | HIS | JAK3 | 08-046 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| JNK1(MAPK8) | GST | JNK1(MAPK8) | 04-163 | Biotin-Modified Erktide | BTSU03 | 10 | ASBF01 | H | |
| JNK2(MAPK9) | GST | JNK2(MAPK9) | 04-164 | Biotin-Modified Erktide | BTSU03 | 10 | ASBF01 | H | |
| JNK3 | GST | JNK3 | 04-150 | Biotin-Modified Erktide | BTSU03 | 10 | ASBF01 | H | |
| KDR | GST | KDR | 08-191 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| KIT | GST | KIT | 08-156 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| KIT[D816E] | GST | KIT[D816E] | 08-541 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| KIT[D816V] | GST | KIT[D816V] | 08-505 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| KIT[D816Y] | GST | KIT[D816Y] | 08-534 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| KIT[T670I] | GST | KIT[T670I] | 08-195 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| KIT[V560G] | GST | KIT[V560G] | 08-504 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| LATS1／MOBKL1A | GST | LATS1／MOBKL1A | 01-523 | Biotin-SGKtide | BTSU15 | 10 | ASBF01 | L | |
| LATS2／MOBKL1A | GST | LATS2／MOBKL1A | 01-524 | Biotin-SGKtide | BTSU15 | 10 | ASBF01 | L | |
| LCK | GST | LCK | 08-170 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| LOK(STK10) | GST | LOK(STK10) | 07-115 | Biotin-Moesin-derived peptide | BTSU18 | 10 | ASBF01 | L | |
| LTK | GST | LTK | 08-106 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| LYNa | GST | LYNa | 08-171 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| LYNb | GST | LYNb | 08-172 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| MAP4K2 | GST | MAP4K2 | 07-111 | Biotin-S6K2 peptide | BTSU14 | 10 | ASBF01 | L | |
| MAP4K5(KHS1) | GST | MAP4K5(KHS1) | 07-113 | Biotin-S6K2 peptide | BTSU14 | 10 | ASBF01 | L | |
| MAPKAPK2 | GST | MAPKAPK2 | 02-142 | Biotin-GS peptide | BTSU05 | 10 | ASBF01 | L | |
| MAPKAPK3 | GST | MAPKAPK3 | 02-143 | Biotin-GS peptide | BTSU05 | 10 | ASBF01 | L | |
| MAPKAPK5 | GST | MAPKAPK5 | 02-144 | Biotin-GS peptide | BTSU05 | 10 | ASBF01 | L | |
| MARK1 | GST | MARK1 | 02-120 | Biotin-CHKtide | BTSU08 | 10 | ASBF01 | L | |
| MARK2 | GST | MARK2 | 02-121 | Biotin-CHKtide | BTSU08 | 10 | ASBF01 | L | |
| MARK3 | GST | MARK3 | 02-122 | Biotin-CHKtide | BTSU08 | 10 | ASBF01 | L | |
| MARK4 | GST | MARK4 | 02-123 | Biotin-CHKtide | BTSU08 | 10 | ASBF01 | L | |
| MELK | GST | MELK | 02-124 | Biotin-GS peptide | BTSU05 | 50 | ASBF01 | L | |
| MER(MERTK) | GST | MER(MERTK) | 08-108 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| MET | GST | MET | 08-151 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| MET[D1228H] | GST | MET[D1228H] | 08-540 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| MET[M1250T] | GST | MET[M1250T] | 08-545 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| MET[Y1235D] | GST | MET[Y1235D] | 08-198 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| MINK | GST | MINK | 07-139 | Biotin-Modified Erktide | BTSU03 | 10 | ASBF01 | H | |
| MNK1(MKNK1) | GST | MNK1(MKNK1) | 02-145 | Biotin-SRPKtide | BTSU70 | 10 | ASBF01 | L | |
| MNK2 | GST | MNK2 | 02-146 | Biotin-SRPKtide | BTSU70 | 10 | ASBF01 | L | |
| MRCKα | GST | MRCKα | 01-107 | Biotin-DAPK1tide | BTSU17 | 50 | ASBF01 | M | |
| MRCKβ | GST | MRCKβ | 01-108 | Biotin-DAPK1tide | BTSU17 | 10 | ASBF01 | L | |
| MSK1 | GST | MSK1 | 01-147 | Biotin-Crosstide | BTSU19 | 10 | ASBF01 | L | |
| MSK2 | GST | MSK2 | 01-148 | Biotin-Crosstide | BTSU19 | 50 | ASBF01 | L | |
| MSSK1 | GST | MSSK1 | 04-159 | Biotin-DYRKtide | BTSU06 | 10 | ASBF01 | L | |
| MST1 | GST | MST1 | 07-116 | Biotin-IRS1 | BTSU07 | 50 | ASBF01 | L | |
| MST2(STK3) | GST | MST2(STK3) | 07-117 | Biotin-IRS1 | BTSU07 | 50 | ASBF01 | L | |
| MST3 | GST | MST3 | 07-118 | Biotin-Moesin-derived peptide | BTSU18 | 10 | ASBF01 | L | |
| MST4 | GST | MST4 | 07-119 | Biotin-Moesin-derived peptide | BTSU18 | 10 | ASBF01 | L | |
| MUSK | GST | MUSK | 08-153 | Biotin-CSKtide | BTSU02 | 50 | ASBF01 | L | |
| NDR1 | GST | NDR1 | 01-125 | Biotin-SGKtide | BTSU15 | 10 | ASBF01 | L | |
| NDR2 | GST | NDR2 | 01-126 | Biotin-SGKtide | BTSU15 | 10 | ASBF01 | L | |
| NEK1 | GST | NEK1 | 05-123 | Biotin-CDK7 peptide | BTSU11 | 10 | ASBF01 | L | |
| NEK2 | Tag-free | NEK2 | 05-226 | Biotin-CDK7 peptide | BTSU11 | 50 | ASBF01 | L | |
| NEK4 | GST | NEK4 | 05-128 | Biotin-GS peptide | BTSU05 | 10 | ASBF01 | L | |
| NEK6 | GST | NEK6 | 05-130 | Biotin-CDK7 peptide | BTSU11 | 50 | ASBF01 | L | |
| NEK7 | GST | NEK7 | 05-131 | Biotin-CDK7 peptide | BTSU11 | 50 | ASBF01 | M | |
| NEK9 | GST | NEK9 | 05-133 | Biotin-CDK7 peptide | BTSU11 | 10 | ASBF01 | L | |
| NIM1K(MGC42105) | GST | NIM1K(MGC42105) | 02-175 | Biotin-CHKtide | BTSU08 | 10 | ASBF01 | L | |
| NPM1-ALK | GST | NPM1-ALK | 08-517 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| NuaK1 | GST | NuaK1 | 02-126 | Biotin-CHKtide | BTSU08 | 10 | ASBF01 | L | |
| NuaK2 | GST | NuaK2 | 02-127 | Biotin-CHKtide | BTSU08 | 10 | ASBF01 | L | |
| p38α(MAPK14) | GST | p38α(MAPK14) | 04-152 | Biotin-Modified Erktide | BTSU03 | 10 | ASBF01 | L | |
| p38β | GST | p38β | 04-153 | Biotin-Modified Erktide | BTSU03 | 10 | ASBF01 | L | |
| p38γ | GST | p38γ | 04-155 | Biotin-Modified Erktide | BTSU03 | 10 | ASBF01 | L | |
| p38δ | GST | p38δ | 04-154 | Biotin-Modified Erktide | BTSU03 | 50 | ASBF01 | L | |
| p70S6K(RPS6KB1) | GST | p70S6K(RPS6KB1) | 01-156 | Biotin-S6K2 peptide | BTSU14 | 10 | ASBF01 | L | |
| p70S6Kβ(RPS6KB2) | GST | p70S6Kβ(RPS6KB2) | 01-155 | Biotin-S6K2 peptide | BTSU14 | 10 | ASBF01 | L | |
| PAK1 | GST | PAK1 | 07-123 | Biotin-LIMKtide | BTSU24 | 10 | ASBF01 | L | |
| PAK2 | GST | PAK2 | 07-124 | Biotin-DAPK1tide | BTSU17 | 10 | ASBF01 | L | |
| PAK4 | GST | PAK4 | 07-126 | Biotin-SGKtide | BTSU15 | 10 | ASBF01 | L | |
| PAK5 | GST | PAK5 | 07-127 | Biotin-DAPK1tide | BTSU17 | 10 | ASBF01 | L | |
| PAK6 | GST | PAK6 | 07-128 | Biotin-SGKtide | BTSU15 | 10 | ASBF01 | L | |
| PASK | GST | PASK | 02-128 | Biotin-GS peptide | BTSU05 | 10 | ASBF01 | L | |
| PBK | GST | PBK | 05-168 | Biotin-Histone H3 peptide | BTSU23 | 10 | ASBF01 | L | |
| PDGFRα(PDGFRA) | GST | PDGFRα(PDGFRA) | 08-157 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| PDGFRα(PDGFRA)[D842V] | GST | PDGFRα(PDGFRA)[D842V] | 08-506 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| PDGFRα(PDGFRA)[T674I] | GST | PDGFRα(PDGFRA)[T674I] | 08-503 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| PDGFRα(PDGFRA)[V561D] | GST | PDGFRα(PDGFRA)[V561D] | 08-507 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| PDGFRβ(PDGFRB) | GST | PDGFRβ(PDGFRB) | 08-158 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| PDHK2 | GST | PDHK2 | 10-140 | Biotin-PDHKtide | BTSU35 | 50 | ASBF01 | L | |
| PDHK4 | GST | PDHK4 | 10-125 | Biotin-PDHKtide | BTSU35 | 50 | ASBF01 | L | |
| PDK1(PDPK1) | GST | PDK1(PDPK1) | 01-132 | Biotin-T308tide | BTSU38 | 10 | ASBF01 | M | |
| PGK | GST | PGK | 01-142 | Biotin-Kemptide | BTSU09 | 10 | ASBF01 | L | |
| PHKG1 | GST | PHKG1 | 02-152 | Biotin-GS peptide | BTSU05 | 10 | ASBF01 | L | |
| PHKG2 | GST | PHKG2 | 02-153 | Biotin-GS peptide | BTSU05 | 10 | ASBF01 | L | |
| PIM1 | HIS | PIM1 | 02-054 | Biotin-S6K2 peptide | BTSU14 | 10 | ASBF01 | L | |
| PIM2 | GST | PIM2 | 02-155 | Biotin-S6K2 peptide | BTSU14 | 10 | ASBF01 | L | |
| PIM3 | GST | PIM3 | 02-156 | Biotin-S6K2 peptide | BTSU14 | 10 | ASBF01 | L | |
| PKACα(PRKACA) | GST | PKACα(PRKACA) | 01-127 | Biotin-Kemptide | BTSU09 | 10 | ASBF01 | L | |
| PKACβ(PRKACB) | GST | PKACβ(PRKACB) | 01-128 | Biotin-Kemptide | BTSU09 | 10 | ASBF01 | L | |
| PKACγ | GST | PKACγ | 01-129 | Biotin-Kemptide | BTSU09 | 10 | ASBF01 | L | |
| PKCα(PRKCA) | GST | PKCα(PRKCA) | 01-133 | Biotin-PKC peptide | BTSU10 | 10 | ASBF01 | L | |
| PKCβ1(PRKCB1) | GST | PKCβ1(PRKCB1) | 01-134 | Biotin-PKC peptide | BTSU10 | 10 | ASBF01 | L | |
| PKCβ2(PRKCB2) | GST | PKCβ2(PRKCB2) | 01-165 | Biotin-PKC peptide | BTSU10 | 10 | ASBF01 | L | |
| PKCγ(PRKCG) | GST | PKCγ(PRKCG) | 01-137 | Biotin-PKC peptide | BTSU10 | 10 | ASBF01 | L | |
| PKCδ(PRKCD) | GST | PKCδ(PRKCD) | 01-135 | Biotin-PKC peptide | BTSU10 | 10 | ASBF01 | L | |
| PKCε(PRKCE) | GST | PKCε(PRKCE) | 01-136 | Biotin-PKC peptide | BTSU10 | 10 | ASBF01 | L | |
| PKCζ | GST | PKCζ | 01-141 | Biotin-PKC peptide | BTSU10 | 10 | ASBF01 | L | |
| PKCη | GST | PKCη | 01-138 | Biotin-PKC peptide | BTSU10 | 10 | ASBF01 | L | |
| PKCθ(PRKCQ) | GST | PKCθ(PRKCQ) | 01-140 | Biotin-PKC peptide | BTSU10 | 10 | ASBF01 | L | |
| PKCι | GST | PKCι | 01-139 | Biotin-PKC peptide | BTSU10 | 10 | ASBF01 | L | |
| PKD1 | GST | PKD1 | 02-157 | Biotin-GS peptide | BTSU05 | 10 | ASBF01 | L | |
| PKD2 | GST | PKD2 | 02-158 | Biotin-GS peptide | BTSU05 | 10 | ASBF01 | L | |
| PKD3 | GST | PKD3 | 02-159 | Biotin-GS peptide | BTSU05 | 10 | ASBF01 | L | |
| PLK1 | GST | PLK1 | 05-157 | Biotin-CDC25ctide | BTSU21 | 50 | ASBF01 | L | |
| PLK3 | GST | PLK3 | 05-159 | Biotin-CDC25ctide | BTSU21 | 50 | ASBF01 | M | |
| PRKX | GST | PRKX | 01-130 | Biotin-Kemptide | BTSU09 | 50 | ASBF01 | L | |
| PYK2(PTK2B) | GST | PYK2(PTK2B) | 08-138 | Biotin-Blk/Lyntide | BTSU04 | 10 | ASBF01 | L | |
| QIK | GST | QIK | 02-129 | Biotin-AMARA peptide | BTSU28 | 50 | ASBF01 | L | |
| RET | GST | RET | 08-159 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| RET[G691S] | GST | RET[G691S] | 08-522 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| RET[M918T] | GST | RET[M918T] | 08-508 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| RET[S891A] | GST | RET[S891A] | 08-523 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| RET[Y791F] | GST | RET[Y791F] | 08-521 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| ROCK1 | GST | ROCK1 | 01-109 | Biotin-LIMKtide | BTSU24 | 10 | ASBF01 | L | |
| ROCK2 | GST | ROCK2 | 01-110 | Biotin-LIMKtide | BTSU24 | 10 | ASBF01 | L | |
| RON | GST | RON | 08-152 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| ROS(ROS1) | GST | ROS(ROS1) | 08-163 | Biotin-IRS1 | BTSU07 | 10 | ASBF01 | L | |
| RSK1(RPS6KA1) | GST | RSK1(RPS6KA1) | 01-149 | Biotin-S6K peptide | BTSU20 | 10 | ASBF01 | L | |
| RSK2(RPS6KA3) | GST | RSK2(RPS6KA3) | 01-150 | Biotin-S6K peptide | BTSU20 | 10 | ASBF01 | L | |
| RSK3(RPS6KA2) | GST | RSK3(RPS6KA2) | 01-151 | Biotin-S6K peptide | BTSU20 | 10 | ASBF01 | L | |
| RSK4(RPS6KA6) | GST | RSK4(RPS6KA6) | 01-152 | Biotin-S6K peptide | BTSU20 | 10 | ASBF01 | L | |
| SGK | GST | SGK | 01-158 | Biotin-SGKtide | BTSU15 | 10 | ASBF01 | L | |
| SGK2 | GST | SGK2 | 01-159 | Biotin-SGKtide | BTSU15 | 10 | ASBF01 | L | |
| SGK3 | GST | SGK3 | 01-160 | Biotin-SGKtide | BTSU15 | 10 | ASBF01 | L | |
| SIK | GST | SIK | 02-131 | Biotin-AMARA peptide | BTSU28 | 50 | ASBF01 | L | |
| SIK3(QSK) | GST | SIK3(QSK) | 02-130 | Biotin-AMARA peptide | BTSU28 | 50 | ASBF01 | L | |
| SLK | GST | SLK | 07-129 | Biotin-Moesin-derived peptide | BTSU18 | 10 | ASBF01 | L | |
| SRC | GST | SRC | 08-173 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| SRM | GST | SRM | 08-174 | Biotin-Blk/Lyntide | BTSU04 | 10 | ASBF01 | L | |
| SRPK2 | GST | SRPK2 | 04-161 | Biotin-DYRKtide | BTSU06 | 10 | ASBF01 | L | |
| SYK | GST | SYK | 08-176 | Biotin-Blk/Lyntide | BTSU04 | 10 | ASBF01 | L | |
| TAK1-TAB1(MAP3K7) | HIS | TAK1-TAB1(MAP3K7) | 09-019 | Biotin-LRRKtide | BTSU48 | 10 | ASBF01 | L | |
| TAOK2 | GST | TAOK2 | 07-133 | Biotin-TAOKtide | BTSU46 | 50 | ASBF01 | H | |
| TBK1 | GST | TBK1 | 05-115 | Biotin-CKtide | BTSU12 | 10 | ASBF01 | L | |
| TEC | GST | TEC | 08-182 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| TIE2(TEK) | GST | TIE2(TEK) | 08-185 | Biotin-Blk/Lyntide | BTSU04 | 10 | ASBF01 | L | |
| TNIK | GST | TNIK | 07-138 | Biotin-Moesin-derived peptide | BTSU18 | 10 | ASBF01 | L | |
| TNK1 | GST | TNK1 | 08-104 | Biotin-CSKtide | BTSU02 | 50 | ASBF01 | H | |
| TRKA(NTRK1) | GST | TRKA(NTRK1) | 08-186 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| TRKB(NTRK2) | GST | TRKB(NTRK2) | 08-187 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| TRKC(NTRK3) | GST | TRKC(NTRK3) | 08-197 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| TSSK1 | Tag-free | TSSK1 | 02-364 | Biotin-GS peptide | BTSU05 | 10 | ASBF01 | L | |
| TSSK2 | GST | TSSK2 | 02-165 | Biotin-GS peptide | BTSU05 | 50 | ASBF01 | L | |
| TSSK3 | GST | TSSK3 | 02-166 | Biotin-GS peptide | BTSU05 | 10 | ASBF01 | L | |
| TXK | GST | TXK | 08-183 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| TYK2 | GST | TYK2 | 08-147 | Biotin-Srctide | BTSU01 | 50 | ASBF01 | L | |
| TYRO3 | GST | TYRO3 | 08-109 | Biotin-CSKtide | BTSU02 | 10 | ASBF01 | L | |
| WNK1 | GST | WNK1 | 05-179 | Biotin-SPAKtide | BTSU27 | 50 | ASBF01 | H | |
| WNK2 | GST | WNK2 | 05-180 | Biotin-SPAKtide | BTSU27 | 50 | ASBF01 | H | |
| WNK3 | GST | WNK3 | 05-181 | Biotin-SPAKtide | BTSU27 | 50 | ASBF01 | H | |
| WNK4 | BTN | BTN-WNK4 | 05-482-20N | Biotin-SPAKtide | BTSU27 | 50 | ASBF01 | L | |
| GST | WNK4 | 05-182 | Biotin-SPAKtide | BTSU27 | 50 | ASBF01 | H | ||
| YES(YES1) | GST | YES(YES1) | 08-175 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| YES(YES1)[T348I] | GST | YES(YES1)[T348I] | 08-533 | Biotin-Srctide | BTSU01 | 10 | ASBF01 | L | |
| ZAP70 | GST | ZAP70 | 08-177 | Biotin-Blk/Lyntide | BTSU04 | 10 | ASBF01 | L | |
当社ADP-Glo™アッセイプロトコール
- アッセイバッファー（20 mM HEPES、0.01% Triton X-100、 2 mM DTT、pH 7.5）を調製。
- アッセイバッファーを用いて4倍濃度のATP溶液、各キナーゼ反応に必要な添加物を含む4倍濃度の基質/金属溶液と4倍濃度のキナーゼ溶液を調製。
- アッセイバッファー 5µL、4倍濃度のATP溶液 5µL、4倍濃度の基質/金属溶液 5µLおよび4倍濃度のキナーゼ溶液 5µLをポリスチレン製384ウェル黒色マイクロプレート（Corning #3573）の各ウェルで混合し、室温で1時間インキュベート。
- 各ウェルにADP-Glo™試薬（Promega）20 µLを加え、40分以上インキュベート（キナーゼ反応の終了、残存ATPの分解）。
- 各ウェルにKinase Detection Reagent（Promega）40 µLを加え、40分以上インキュベート（ADPをATPに変換）。
- 各ウェルの発光強度を測定（EnVision 2104 (Revvity））し、発光強度を生成ADP濃度へ換算することでキナーゼ活性を評価。
(注）詳細プロトコールはPromega社提供のADP-Glo™ Kinase Assay Technical Manual をご参照ください。
ATPase活性確認の重要性
ATPaseはATPをADPと無機リン酸に加水分解する酵素ですが、in vitroにおいて、キナーゼの中には高い内因性ATPase活性を有しているものが存在しており、そのようなキナーゼを用いたアッセイでは基質非存在下でもATPase活性によりADPが生成されます。ADP-Glo™ Kinase Assayでは、アッセイ中に生成されたADPをATPに戻し、それをルシフェラーゼ反応によって発光として測定します。この発光の量は、キナーゼの活性を示すとされていますが、キナーゼがATPase活性を持っている場合、基質非存在下でもADPが生成されてしまうため、測定結果に影響を与える可能性があります。そのため、このアッセイを使う際には、キナーゼのATPase活性を考慮してデータを取得することが重要です。
カルナでADP-Glo™ Kinase Assayデータ取得済みの各キナーゼにおけるATPase の影響は上記表内 ATPase Activity欄にある強度をご参照ください。
JNK2(MAPK9)をK252aを基質としてADP-Glo™ Kinase Assayで測定
Substrate(-)でもキナーゼ濃度依存的にRLUが上昇。Substrate(+)でもRLUが上昇するが、測定されたRLUの大部分はATPを分解するATPase活性由来であり、キナーゼが基質をリン酸化する特異活性よりもATPase活性の方が強い。
Substrate(-)でもキナーゼ濃度依存的にRLUが上昇。Substrate(+)でもRLUが上昇するが、測定されたRLUの大部分はATPを分解するATPase活性由来であり、キナーゼが基質をリン酸化する特異活性よりもATPase活性の方が強い。
ADP-Glo™以外の方法でキナーゼ活性を測定する場合でも、ATPase活性が強いキナーゼを用いると、反応系のATPが分解されてしまい、結果に影響を与える可能性があります。そのため、こうしたキナーゼについては、ATPの分解による影響を補うために、Mobility Shift Assayなど基質のリン酸化を直接検出するアッセイを併用するのが望ましいです。
JNK2（MAPK9）Mobility Shift Assay測定
Km値付近のATP濃度 : 50 µmol/LではATPaseによるATP分解により特異活性シグナルが頭打ちとなり酵素濃度依存的な基質のリン酸化が測定できなくなる。この様なケースではシグナルが頭打ちとならない酵素濃度設定による測定が必要。
Km値付近のATP濃度 : 50 µmol/LではATPaseによるATP分解により特異活性シグナルが頭打ちとなり酵素濃度依存的な基質のリン酸化が測定できなくなる。この様なケースではシグナルが頭打ちとならない酵素濃度設定による測定が必要。
- カルナ提供 ADP-Glo™アッセイキットの購入、ADP-Glo™アッセイ試験の外部委託をご検討されている方へ
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