Perkin Elmer HTS High Throughput Screening Perkin Elmer

Perkin. Elmer에서 제공하는 고효율 효능 검색 방법 (HTS, High Throughput Screening)

Perkin. Elmer HTS 관련 장비 Performance ”HTS” En. Vision Xcite/HTS ”Research” VICTOR 3 V FUSION VICTOR 3 VICTOR Light Price

Ø Alpha. Screen Ø DELFIA Ø LANCE, Tru. Point

Alpha. Screen 원리

Direct vs Indirect capture biotin-antigen primary Ab • preferred format / most efficient • simple stoichiometry • good control on binding events secondary Ab (or prot. A, G, LA) • • no custom bead conjugation required save expensive primary Ab additional equilibrium introduced needs high affinity primary Ab , selective secondary Ab or Proteins A, G, L, LA

Alpha. Screen 제품 종류 Tag Capture • c-myc • Digoxygenin • FITC • FLAG M 2 • GST • HA • 6 His • Ni 2+ chelate Functional GPCR c. AMP IP 3 Ig. G Capture • g. Ig. G • h. Ig. G • m. Ig. G • r. Ig. G • prot. A p. Y Capture • PY 20 • PT 66 • p. Tyr-100 Conjugation kit 측정 장비 • FUSION ALPHA

DELFIA Dissociation: 형광 물질 분리 Enhanced: 신호 강화 및 증폭 Lanthanide: Lanthanide계열의 Fluoro. Immuno: 형광 물질 Assay: 물질 탐색 방법

Four Lanthanide Metals Samarium, Europium, Terbium, Dysprosium

DELFIA® Assay 원리 Dissociation Enhanced Lanthanide FIA Eu Eu Fluorescence based heterogeneous detection technology utilizing time resolved fluorometry

Time-Resolution Image Prompt signal Time resolved signal Low background TRF microimaging

Conversion from ELISA to DELFIA ü ü ü 보다 넓은 detection 범위 보다 향상된 민감도 시약 절감 간소화된 실험 단계 24시간 이상 안정된 시약의 활성 필요한 시약 및 소모품 - Coated plate/Acrowell filter plate - DELFIA labeled biomolecule - Enhancement solution 선택 시약 - DELFIA Wash Buffer - DELFIA Assay Buffer 측정 장비: VICTOR, FUSION, En. Vision, View. Lux

LACNE & Tru. Point

LANCE 원리 340 nm Binding Eu APC 665 nm Acceptor label: Surelight. TM APC Donor label: Europium chelates • lyophilized - user friendly format • over 15 years of experience • industry leader • patent protected TR-FRET의 원리를 기본적으로 이용

LANCE™ 제품 종류 DONOR LABELS Reagents for kinase assays § Eu-labelled PY 20 § Eu-labelled PT 66 § Eu-labelled PTyr 100 § 11 different Ser/Thr specific antibodies § LANCE kinase start up package For demos Anti-tags antibodies § Eu-labelled anti-GST § Eu-labelled anti-FLAG § Eu-labelled anti-HA § Eu-labelled anti-c-myc § Eu-labelled anti-6*his DETECTION BUFFER § Tris based buffer with BSA Anti-species antibodies § Eu-labelled anti-mouse Ig. G § Eu-labelled anti-rabbit Ig. G § Eu-labelled anti-human Ig. G Generic reagents § Eu-labelled Steptavidin § Eu-labelled Protein G Labelling reagents § Eu & Tb chelates ACCEPTOR LABELS: • • APC-Streptavidin APC-anti-c. Myc APC-anti-FLAG APC-anti-GST APC-anti-6 HIS APC-anti-mouse APC-anti-rabbit APC-WGA

LANCE Tyrosine Kinase Assay LANCE tyrosine kinase reagents • • • Start-up kit package Eu-labeled PT 66 Eu-labelled PY 20 Eu-labelled P-tyr-100 Sure. Light. TM APC acceptors Eu-labeled anti-species antibodies Labeling Service Biazzo-Ashnault et al. (2001) Detection of insulin receptor tyrosine kinase activity using time-resolved fluorescence energy transfer technology. Anal. Biochem. 291, 155 -158

Tru. Point ™ Eu Dye 615 nm Caspase 3 Dye Eu

Tru. Point ™ 제품 종류 • Caspase-3, caspase-6 and caspase-8 (384 -well kits) • Helicase • Caspase-3 (96 -well kit) • Labeling service § Caspase-2 § Caspase-9

Tru. Point 특장점 • Homogenous • 높은 민감도 - Time resolution • Emission at 615 nm, red-shifted – higher quality hits • High quenching efficiency, high S/B ratios • 형광 측정의 안정된 결과값 • 검색단위 최소화 가능으로 인한 시료 절감

고효율 효능 검색 방법 비교 Tyrosine Kinase Assays Leitner et al. Lab. Automation, 1998

Alpha. Screen DELFIA LANCE 측정장비 FUSIONALPHA FUSION, VICTOR, En. Vision, Viwe. Lux 에너지원 680 nm Laser Xenon flash lamp 두 종류의 bead 필요 (Donor, Acceptor) 한 종류의 형광 물질 필요 두 종류의 형광물질 필 요 (Donor, Acceptor) 실험단계 Homogeneous Heterogeneous Homogeneous 자동화 Yes, but contains wash Yes 단백질 결합 측정 범위 p. M to m. M p. M to n. M Plate Type White opaque SA coated clear, white, yellow Black, white, clear Application c. AMP assays Kinase assays Protein binding assays Protease assays Immunoassays Receptor-ligand biding assays Protein-protein binding assays Cytokine assays Adherent cell assays Enzyme assays DNA hybridization assays Cytotoxicity assays Immunoassays Kinase assays Receptor assays Protein binding assays Protease assays Helicasae assay