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MicroBeta. Page 2 MicroBeta Page 3 MicroBeta Page 4 MicroBeta.

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Presentation on theme: "MicroBeta. Page 2 MicroBeta Page 3 MicroBeta Page 4 MicroBeta."— Presentation transcript:

1 MicroBeta

2 Page 2 MicroBeta

3 Page 3 MicroBeta

4 Page 4 MicroBeta

5 Page 5 Liquid Coated plate Filter MicroBeta

6 Page 6 Eppendorf & 4ml 45 x 15 mm

7 Page 7 Rigid or flexible microplates

8 Page 8

9 Page 9 Filtermat

10 Page 10 FilterMate Waste bottles and stands Pressurized wash bottle assembly UniFilter plate in position for harvesting Holder for self-aligning and plain, non-self- aligning filter mats

11 Page 11 Unifilter

12 Page 12 MicroBeta workstation

13 Page 13

14 Page 14

15 Page 15 MicroBeta LSCSPA LuminescenceFilter plates

16 Page 16 MicroBeta

17 Page 17 Normalization and Standardization Protocols NormalizationStandardization

18 Page 18 solventbeta decay fluorlight physicalchemicalcolour Quenching

19 Page 19 Quench Curve

20 Page 20

21 Page 21

22 Page 22 BIBW2992, an irreversible EGFR/HER2 inhibitor highly effective in preclinical lung cancer models Oncogene, April 14, 2008

23 Page 23 33 P[ATP] In vitro kinase activity assay For VEGF2 and HGFR kinase assays, a random polymer pEY (4:1) from SIGMA was used as substrate. Each 50 µl-reaction contained 5 % Me2SO, 40 mM HEPES pH 7.4, 5 mM MgCl2 0.5 mg/ml pEY, 0.05% Triton X-100, 100 µM ATP, 1 µCi [γ 33P] ATP and 10 µl of enzyme preparation. … … The precipitate was then trapped onto GF/B Unifilters (Packard, 6005177) using a 96 well filter mate universal harvester (Packard, C961961). … … radioactivity was determined by scintillation counting using a TopCount (Packard) or a Microbeta b counter (Wallac)

24 Page 24 Scintillation Proximity Assay

25 Page 25 Scintillation Proximity Assay

26 Page 26 White plastic Coating of scintillator Flashplate

27 Page 27 FlashPlate Scintillation Proximity Assays for Characterization and Screening of DNA Polymerase, Primase, and Helicase Activities J Biomol Screen.2001; 6: 39-46 FlashPlate Scintillation Proximity Assays

28 Page 28 MAP (mitogen activated protein) kinase activity can be measured through the use of MBP (myelin basic protein) as a substrate in the presence of appropriate buffer and [33P]-g-ATP. Activated MAP kinase incorporates the 33P onto the serine and threonine residues of MBP. When increasing amounts of MBP are coated onto FlashPlate, counts bound increase without increasing background (Fig. 1). Flashplate MAP Kinase

29 Page 29 Luminescence Single photon event Easier to remove background High speed electronics required up to 20 x 10 6 CPS Temperature control option for MicroBeta X …have optimised luminescence performance

30 Page 30 Any Questions? Thank you!


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