TECHNICAL RESOURCES

PerkinElmer단시간에 단백질 정량까지 가능한 Kinase assay kit

2018-12-26 09:12



하루 종일 걸리는 western blot은 이제 그만!

Kinase protein의 정확한 발현량 확인까지 가능한 PerkinElmer의 Alpha SureFire Cellular Kinase Assay Kits


단백질 loading 양이 적어 질수록 미약하게 보이는 밴드 때문에 고생이셨나요?

하루종일 시간 투자를 해서 얻은 결과가 만족스럽지 않아서 다시 실험을 진행하셔야 했나요?

PerkinElmer의 AlphaScreen® SureFire®, AlphaLISA® SureFire® Ultra, Alpha SureFire ultra Multiplex를 사용하여 드라마틱한 결과를 확인해 보세요.














[Fig 1.] 좌) Western blot, 우) AlphaScreen® SureFire® Kit 결과 비교


1) Alpha SureFire® Cellcular Kinase Detection Kit


  • Alpha Donor bead와 acceptor bead 간의 거리가 phospholyated protein (analyte)에 의해 가까워지면, excitation 된 donor bead로부터 나온 에너지가 acceptor bead로 전달되어 방출되는 emission 값을 측정하는 원리입니다.
  • Sandwich immunoassays
  • Western blot과 달리 cell에 존재하는 phosphorylated protein detection을 정량할 수 있습니다.

  • Washing step이 없고, 전체 실험 과정이 간단합니다.

  • 자동화 시스템에 적용할 수 있습니다.

  • 세 가지 format의 kit

      AlphaScreen® SureFire®

    • 2005년부터 사용되어 온 original format
    • Protein A가 conjugation된 acceptor bead를 사용
    • 4시간 이내 실험 완료
    • Kit 이외에 Protein A를 detection 할 수 있는 AlphaScreen 
                Protein A general IgG kit 구입 필수







      AlphaLISA® SureFire® Ultra

    • CaptSure-antibody가 coating 되어진 acceptor bead가 analyte를 인식하는 antibody에 conjugation된 CapSure™을 인식하는 format
    • 낮은 백그라운드 signal, 향상된 sensitivity와 dynamic range
    • 관련 없는 antibody로부터는 signal 방해를 안 받기 때문에 항체 신약 screening에 이상적
    • Hemoglobin이 포함된 샘플에 최적화된 AlphaLISA bead
    • 2시간 이내 실험 완료
    • Kit 이외에 추가 구매 시약 없음




      Alpha SureFire® Ultra multiplex

    • 동일한 well에서 multiple signal을 detection
    • 동일한 well에서 internal control (e.g., total protein)을 함께 사용함으로써 실험의 퀄리티가 향상되며, 시간과 비용을 절감
    • AlphaLISA® SureFire® Ultra™와 같이 별도 추가 구매할 시약 없음













2) 실험 방법

* 384 well white plate 기준


















3) Product List

Target
AlphaScreen SureFire
AlphaLISA SureFire Ultra
500 assay
10,000 assay
50,000 assay
100 assay
500 assay
10,000 assay
50,000 assay
p-4EBP1 (Thr37/46)
TGR4ES500
TGR4ES10K
TGR4ES50K
ALSU-P4EBP-A-HV
ALSU-P4EBP-A500
ALSU-P4EBP-A10K
ALSU-P4EBP-A50K
p-4EBP1 (Thr70)
TGR4E2S500
TGR4E2S10K
TGR4E2S50K
 
 
 
 
p-ACC (Ser79)
 
 
 
ALSU-PACC-A-HV
ALSU-PACC-A500
ALSU-PACC-A510K
ALSU-PACC-A50K
p-AKT1 (Thr308)
TGRA2S500
TGRA2S10K
TGRA2S50K
 
 
 
 
p-AKT1 (Ser473)
TGRA5S500
TGRA5S10K
TGRA5S50K
 
 
 
 
Total AKT1
TGRTA1S500
TGRTA1S10K
TGRTA1S50K
ALSU-TAKT1-A-HV
ALSU-TAKT1-A500
ALSU-TAKT1-A10K
ALSU-TAKT1-A50K
p-AKT1/2/3 (Thr308)
TGRA3S500
TGRA3S10K
TGRA3S50K
ALSU-PAKT-A-HV
ALSU-PAKT-A500
ALSU-PAKT-A10K
ALSU-PAKT-A50K
p-AKT1/2/(3) (Ser473)
TGRAS500
TGRAS10K
TGRAS50K
 
 
 
 
p-AKT1/2/3 (Ser473)
TGRA4S500
TGRA4S10K
TGRA4S50K
ALSU-PAKT-B-HV
ALSU-PAKT-B500
ALSU-PAKT-B10K
ALSU-PAKT-B50K
Total AKT1/2/3
TGRTAPS500
TGRTAPS10K
TGRTAPS50K
 
 
 
 
p-ALK (Tyr1586)
TGRALS500
TGRALS10K
TGRALS50K
 
 
 
 
p-ALK (Tyr1604)
TGRAL2S500
TGRAL2S10K
TGRAL2S50K
 
 
 
 
Total ALK
TGRTALS500
TGRTALS10K
TGRTALS50K
 
 
 
 
p-AMPKα (Thr172)
 
 
 
ALSU-PAMPK-A-HV
ALSU-PAMPK-A500
ALSU-PAMPKA-10K
ALSU-PAMPKA-50K
p-ATF-1 (Ser63)
TGRAF1S500
TGRAF1S10K
TGRAF1S50K
 
 
 
 
p-ATF-2 (Thr71)
TGRAF2S500
TGRAF2S10K
TGRAF2S50K
 
 
 
 
P-BAD (Ser112)
TGRBS500
TGRBS10K
TGRBS50K
 
 
 
 
p-BAD (Ser136)
TGRB2S500
TGRB2S10K
TGRB2S50K
 
 
 
 
p-ß-Catenin (Ser45)
TGRBCS500
TGRBCS10K
TGRBCS50K
 
 
 
 
p-Caspase 9 (Ser196)
TGRC9S500
TGRC9S10K
TGRC9S50K
 
 
 
 
p-Chk1 (Ser345)
TGRCHK1S500
TGRCHK1S10K
TGRCHK1S50K
 
 
 
 
p-Chk2 (Thr68)
TGRCHK2S500
TGRCHK2S10K
TGRCHK2S50K
 
 
 
 
p-c-Jun (Ser63)
TGRCJS500
TGRCJS10K
TGRCJS50K
 
 
 
 
p-c-Jun (Ser73)
TGRCJ2S500
TGRCJ2S10K
TGRCJ2S50K
 
 
 
 
p-c-Met (Tyr1234/1235)
TGRCMS500
TGRCMS10K
TGRCMS50K
ALSU-PCMET-A-HV
ALSU-PCMET-A500
ALSU-PCMET-A10K
ALSU-PCMET-A50K
p-Cofilin (Ser3)
TGRCOFS500
TGRCOFS10K
TGRCOFS50K
 
 
 
 
p-CREB-1 (Ser133)
TGRCBS500
TGRCBS10K
TGRCBS50K
ALSU-PCREB-A-HV
ALSU-PCREB-A500
ALSU-PCREB-A10K
ALSU-PCREB-A50K
p-EGF Receptor
(Tyr1068)
TGRERS500
TGRERS10K
TGRERS50K
ALSU-PEGFR-A-HV
ALSU-PEGFR-A500
ALSU-PEGFR-A10K
ALSU-PEGFR-A50K
p-eIF2alpha (Ser51)
 
 
 
ALSU-PEIF2-B-HV
ALSU-PEIF2-B500
ALSU-PEIF2-B10K
ALSU-PEIF2-B50K
p-eIF4E (Ser209)
TGREIF4S500
TGREIF4S10K
TGREIF4S50K
ALSU-PEIF4-A-HV
ALSU-PEIF4-A500
ALSU-PEIF4-A10K
ALSU-PEIF4-A50K
p-Elk-1 (Ser383)
TGRELKS500
TGRELKS10K
TGRELKS50K
 
 
 
 
p-ErbB2 (Tyr1221/1222)
TGREB2S500
TGREB2S10K
TGREB2S50K
ALSU-PEB2-A-HV
ALSU-PEB2-A500
ALSU-PEB2-A10K
ALSU-PEB2-A50K
p-ERK1/2 (T202/Y204)
TGRESB500
TGRESB10K
TGRESB50K
ALSU-PERK-A-HV
ALSU-PERK-A500
ALSU-PERK-A10K
ALSU-PERK-A50K
Total ERK1/2
TGRTESB500
TGRTESB10K
TGRTESB50K
ALSU-TERK-A-HV
ALSU-TERK-A500
ALSU-TERK-A10K
ALSU-TERK-A50K
Total GAPDH (human)
TGRGDS500
TGRGDS10K
TGRGDS50K
ALSU-TGAPD-A-HV
ALSU-TGAPD-A500
ALSU-TGAPDA10K
ALSU-TGAPDA50K
Total GAPDH
(human, mouse, rat)
 
 
 
ALSU-TGAPD-B-HV
ALSU-TGAPD-B500
ALSU-TGAPD-B10K
ALSU-TGAPD-B50K
p-GSK3α (Ser21)
TGRGAS500
TGRGAS10K
TGRGAS50K
 
 
 
 
p-GSK3β (Ser9)
TGRGBS500
TGRGBS10K
TGRGBS50K
ALSU-PGS3B-A-HV
ALSU-PGS3B-A500
ALSU-PGS3B-A10K
ALSU-PGS3B-A50K
p-Histone H3 (Ser10)
TGRH3S500
TGRH3S10K
TGRH3S50K
 
 
 
 
p-HSP27 (Ser82)
TGRHS27S500
TGRHS27S10K
TGRHS27S50K
 
 
 
 
p-IGF1 Receptor β
(Tyr1135/1136)
 
 
 
ALSU-PIGFR-B-HV
ALSU-PIGFR-B500
ALSU-PIGFR-B10K
ALSU-PIGFR-B50K
p-IκBα (Ser32/36)
TGRIKS500
TGRIKS10K
TGRIKS50K
 
 
 
 
Total IκBα
TGRTIKS500
TGRTIKS10K
TGRTIKS50K
 
 
 
 
p-IKKα (Ser176/180)
TGRKAS500
TGRKAS10K
TGRKAS50K
ALSU-PIKKA-A-HV
ALSU-PIKKA-A500
ALSU-PIKKA-A10K
ALSU-PIKKA-A50K
p-IKKβ (Ser177/181)
TGRKBS500
TGRKBS10K
TGRKBS50K
 
 
 
 
p-Insulin Receptor β
(Tyr1150/1151)
TGRIRS500
TGRIRS10K
TGRIRS50K
ALSU-PINR-A-HV
ALSU-PINR-A500
ALSU-PINR-A10K
ALSU-PINR-A50K
p-JNK1/3
(Thr183/Tyr185)
TGRJS500
TGRJS10K
TGRJS50K
 
 
 
 
p-JNK1/2/3
(Thr183/Tyr185)
TGRJPS500
TGRJPS10K
TGRJPS50K
ALSU-PJNK-A-HV
ALSU-PJNK-A500
ALSU-PJNK-A10K
ALSU-PJNK-A50K
Total JNK1/2/3
TGRTJS500
TGRTJS10K
TGRTJS50K
 
 
 
 
p-MEK1 (Ser218/222)
 
 
 
ALSU-PMEK1-A-HV
ALSU-PMEK1-A500
ALSU-PMEK1-A10K
ALSU-PMEK1-A50K
Total MEK1
 
 
 
ALSU-TMEK1-A-HV
ALSU-TMEK1-A500
ALSU-TMEK1-A10K
ALSU-TMEK1-A50K
p-MKK3/6
(Ser189/Ser207)
TGRMK3S500
TGRMK3S10K
TGRMK3S50K
 
 
 
 
p-MKK4
(Ser257/Thr261)
TGRMK4S500
TGRMK4S10K
TGRMK4S50K
 
 
 
 
p-mTOR (Ser2448)
 
 
 
ALSU-PMTOR-A-HV
ALSU-PMTOR-A500
ALSU-PMTOR-A10K
ALSU-PMTOR-A50K
Total mTOR
 
 
 
ALSU-TMTOR-A-HV
ALSU-TMTOR-A500
ALSU-TMTOR-A10K
ALSU-TMTOR-A50K
p-NFκB p65 (Ser536)
TGRNFS500
TGRNFS10K
TGRNFS50K
ALSU-PNFKB-A-HV
ALSU-PNFKB-A500
ALSU-PNFKB-A10K
ALSU-PNFKB-A50K
Total NFkB p65
TGRTNFS500
TGRTNFS10K
TGRTNFS50K
 
 
 
 
P-p38 MAPK
(Thr180/Tyr182)
 
 
 
ALSU-PP38-B-HV
ALSU-PP38-B500
ALSU-PP38-B10K
ALSU-PP38-B50K
Total p38 MAPK
 
 
 
ALSU-TP38-B-HV
ALSU-TP38-B500
ALSU-TP38-B10K
ALSU-TP38-B50K
p-p53 (Ser15)
TGRP53S500
TGRP53S10K
TGRP53S50K
 
 
 
 
p-p53 (Ser392)
TGRP532S500
TGRP532S10K
TGRP532S50K
 
 
 
 
Total p53
 
 
 
 
 
 
 
p-p70S6K (Thr229)
TGR703S500
TGR703S10K
TGR703S50K
 
 
 
 
p-p70S6K (Thr389)
TGR70S500
TGR70S10K
TGR70S50K
ALSU-PP70-A-HV
ALSU-PP70-A500
ALSU-PP70-A10K
ALSU-PP70-A50K
p-p70S6K
(Thr421/Ser424)
TGR702S500
TGR702S10K
TGR702S50K
 
 
 
 
Total p70S6K
TGRT70S500
TGRT70S10K
TGRT70S50K
 
 
 
 
p-PDK1 (Ser241)
TGRPS500
TGRPS10K
TGRPS50K
 
 
 
 
p-S6RP (Ser235/236)
TGRS6PS500
TGRS6PS10K
TGRS6PS50K
 
 
 
 
p-S6RP (Ser240/244)
 
 
TGRS6P2S50K
ALSU-PS6R-A-HV
ALSU-PS6R-A500
ALSU-PS6R-A10K
ALSU-PS6R-A50K
p-SLP76 (Ser376)
 
 
 
ALSU-PSLP-A-HV
ALSU-PSLP-A500
ALSU-PSLP-A10K
ALSU-PSLP-A50K
Total-SLP76
 
 
 
ALSU-TSLP-A-HV
ALSU-TSLP-A500
ALSU-TSLP-A10K
ALSU-TSLP-A50K
p-SMAD1 (Ser463/465)
TGRSM1S500
TGRSM1S10K
TGRSM1S50K
ALSU-PSM1-A-HV
ALSU-PSM1-A500
ALSU-PSM1-A10K
ALSU-PSM1-A50K
p-SMAD2 (Ser465/467)
TGRSM2S500
TGRSM2S10K
TGRSM2S50K
 
 
 
 
p-SMAD3 (Ser423/425)
TGRSM3S500
TGRSM3S10K
TGRSM3S50K
ALSU-PSM3-A-HV
ALSU-PSM3-A500
ALSU-PSM3-A10K
ALSU-PSM3-A50K
p-STAT1 (Tyr701)
TGRS1S500
TGRS1S10K
TGRS1S50K
ALSU-PST1-A-HV
ALSU-PST1-A500
ALSU-PST1-A10K
ALSU-PST1-A50K
p-STAT1 (Ser727)
 
 
 
ALSU-PST1-B-HV
ALSU-PST1-B500
ALSU-PST1-B10K
ALSU-PST1-B50K
p-STAT3 (Tyr705)
TGRS3S500
TGRS3S10K
TGRS3S50K
ALSU-PST3-A-HV
ALSU-PST3-A500
ALSU-PST3-A10K
ALSU-PST3-A50K
Total STAT3
TGRTS3S500
TGRTS3S10K
TGRTS3S50K
ALSU-TST3-A-HV
ALSU-TST3-A500
ALSU-TST3-A10K
ALSU-TST3-A50K
p-STAT4 (Tyr693)
 
 
 
ALSU-PST4-A-HV
ALSU-PST4-A500
ALSU-PST4-A10K
ALSU-PST4-A50K
p-STAT5 (Tyr694/699)
TGRS5S500
TGRS5S10K
TGRS5S50K
ALSU-PST5-B-HV
ALSU-PST5-B500
ALSU-PST5-B10K
ALSU-PST5-B50K
Total STAT5
 
 
 
ALSU-TST5-A-HV
ALSU-TST5-A500
ALSU-TST5-A10K
ALSU-TST5-A50K
p-STAT6 (Tyr641)
 
 
 
ALSU-PST6-A-HV
ALSU-PST6-A500
ALSU-PST6-A10K
ALSU-PST6-A50K
p-SYK (Tyr525/526)
 
 
 
ALSU-PSYK-A-HV
ALSU-PSYK-A500
ALSU-PSYK-A10K
ALSU-PSYK-A50K
Total SYK
 
 
 
ALSU-TSYK-A-HV
ALSU-TSYK-A500
ALSU-TSYK-A10K
ALSU-TSYK-A50K
p-Topoisomerase Iiα
(Ser1106)
TGRT2A2S500
TGRT2A2S10K
TGRT2A2S50K
 
 
 
 
p-Topoisomerase Iiα
(Thr1343)
TGRT2AS500
TGRT2AS10K
TGRT2AS50K
 
 
 
 
Total Topoisomerase IIα
TGRTT2AS500
TGRTT2AS10K
TGRTT2AS50K
 
 
 
 
p-TrkA(Tyr674/675)/
p-TrkB (Tyr706/707)
 
 
 
ALSU-PTRKAB-A-HV
ALSU-PTRKAB-A500
ALSU-PTRKAB-A10K
ALSU-PTRKAB-A50K
p-VEGF Receptor 2
(Tyr1175)
TGRVRS500
TGRVRS10K
TGRVRS50K
ALSU-PVGFR-A-HV
ALSU-PVGFR-A500
ALSU-PVGFR-A10K
ALSU-PVGFR-A50K
Total VEGF Receptor 2
 
 
 
ALSU-TVGFR-A-HV
ALSU-TVGFR-A500
ALSU-TVGFR-A10K
ALSU-TVGFR-A50K
p-AKT 1/2/3 (Ser473) (Eu) + Total AKT1 (Tb)
 
 
 
MPSU-PTAKT-K-HV
MPSU-PTAKT-K500
MPSU-PTAKT-K10K
MPSU-PTAKT-K50K
p-ERK 1/2 (Eu) +
Total ERK (Tb)
 
 
 
MPSU-PTERK-K-HV
MPSU-PTERK-K500
MPSU-PTERK-K10K
MPSU-PTERK-K50K


제품별 자세한 설명은 PerkinElmer 홈페이지에서 확인 가능합니다.


PerkinElmer 제품 홈페이지


제품 문의: T.031-728-3235  E. hj.eo@dawinbiotech.com

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