SERVIZIO BIO-PLEX CRIBI

Presentazione sul tema: "SERVIZIO BIO-PLEX CRIBI"— Transcript della presentazione:

1 SERVIZIO BIO-PLEX CRIBI
Using this method, over 100 distinct microsphere sets can be created. It is important to remember that these beads, or microspheres are suspended in solution, so they are free to interact with the sample. This creates an assay platform that is - flexible (protein, peptides, DNA - reproducible ( as beads are uniformly distributed in solution) - improved sensitivity (better S/N, and near solution-phase kinetics) The object is to conjugate, or couple a different biological reaction to the surface of each bead set

2 Suspension Array reader
Bio Plex A N L I S M U T E Suspension Array reader D I P R O T E N

3 Analisi qualitative e quantitative simultanee di proteine

4 Principio Funzionale Using this method, over 100 distinct microsphere sets can be created. It is important to remember that these beads, or microspheres are suspended in solution, so they are free to interact with the sample. This creates an assay platform that is - flexible (protein, peptides, DNA - reproducible ( as beads are uniformly distributed in solution) - improved sensitivity (better S/N, and near solution-phase kinetics) The object is to conjugate, or couple a different biological reaction to the surface of each bead set

5 Principio Funzionale Using this method, over 100 distinct microsphere sets can be created. It is important to remember that these beads, or microspheres are suspended in solution, so they are free to interact with the sample. This creates an assay platform that is - flexible (protein, peptides, DNA - reproducible ( as beads are uniformly distributed in solution) - improved sensitivity (better S/N, and near solution-phase kinetics) The object is to conjugate, or couple a different biological reaction to the surface of each bead set

6 Principio Funzionale Using this method, over 100 distinct microsphere sets can be created. It is important to remember that these beads, or microspheres are suspended in solution, so they are free to interact with the sample. This creates an assay platform that is - flexible (protein, peptides, DNA - reproducible ( as beads are uniformly distributed in solution) - improved sensitivity (better S/N, and near solution-phase kinetics) The object is to conjugate, or couple a different biological reaction to the surface of each bead set

7 La Camera di Lettura Using this method, over 100 distinct microsphere sets can be created. It is important to remember that these beads, or microspheres are suspended in solution, so they are free to interact with the sample. This creates an assay platform that is - flexible (protein, peptides, DNA - reproducible ( as beads are uniformly distributed in solution) - improved sensitivity (better S/N, and near solution-phase kinetics) The object is to conjugate, or couple a different biological reaction to the surface of each bead set

8 M U L T I W E B I O P L E X Reporter channel Classify channel
To perform a test, thousands of probes, such as antibodies are bound to the microsphere. Reporter channel Classify channel

9 Saggi per Citochine

10 Sandwich Immunoassay - Cytokine -
Saggi per Citochine Sandwich Immunoassay - Cytokine - IL-2 IL-4 TNFa GmCSF

11 Kit applicativi Citochine umane (27): Nuovo kit 27-plex:
Multiplex tested Citochine umane (27): IL-2 IL-1b IL-4 IL-5 IL-6 IL-7 IL-8 IL-12 (p70) IL-10 IL-13 IFN- IL-17 TNF- MCP-1 (MCAF) GM-CSF MIP-1b Eotaxin Rantes IL-1ra IL-9 IL-15 PDGF bFGF VEGF Nuovo kit 27-plex: 171-A HUMAN CYTOKINE 27-PLEX, 1X96

12 Kit applicativi Citochine di topo (23): Nuovo kit 23-plex:
Multiplex tested Citochine di topo (23): IL-1 IL-1a MIP-1a IL-5 IL-3 Rantes IL-2 IL-6 IL-13 IL-10 IL-12 (p40) MCP1 IL-4 IL-12 (p70) Eotaxin IFN- IL-17 MIP-1 TNF- KC IL-9 GM-CSF Nuovo kit 23-plex: 171F MOUSE CYTOKINE 23-PLEX, 1X96

13 Kit applicativi Citochine di Ratto (9): Kit 9-plex:
Multiplex tested Citochine di Ratto (9): GM-CSF IL-4 IFN- IL-6 IL-1 IL-10 IL-2 TNF- Kit 9-plex: 171K11070 RAT CYTOKINES 9-PLEX A PANEL,1X96

14 Saggi per Fosfoproteine

15 Sandwich Immunoassay – Phospho -
Saggi per Fosfoproteine Sandwich Immunoassay – Phospho - JNK IB ERK1 STAT3 P

16 Saggi per Fosfoproteine
15 Phosphoprotein Assays Phospho-Erk2 Phospho-IB- Phospho-JNK Phospho-p38 MAPK Phospho-Stat3 Phospho-ATF2 Phospho-Erk1 Phospho-Erk1/2 Phospho-EGFR Phospho-Akt Phospho-GSK-3a/b Phospho-p70 S6 Phospho-p90 RSK Phospho-Stat2 Phospho-TrkA Phospho C-Jun Phospho HSP 27 Phospho p53 Phospho NF-kβ Phospho S6 Ribosomal P. LISATI DI COLTURE CELLULARI LISATI DI CAMPIONI TISSUTALI

17 Saggi per Fosfoproteine
8 Total Target Assays Total Erk2 Total IB Total p38 MAPK Total ATF-2 Total JNK Total Akt Total p90 RSK Total c-Jun

18 Correlazione tra Bio-Plex e W. Blot
All of our cytokine and phosphoprotein detection assays are validated against ELISA and western blot to validate the results. Here we look at the correlation between the Bio-Plex phosphoprotein detection assay and western blot for Erk and Stat-3. Phospho-Erk Phospho-Stat3 Western Blot, 8 ug protein/lane Western Blot, 8 ug protein/lane Bio-Plex Assay Bio-Plex Assay 1600 1200 MFI of Bio-Plex +2SD 800 400 1 2 3 4 5 6

19 Un Sistema Aperto Formati Applicabili
Using this method, over 100 distinct microsphere sets can be created. It is important to remember that these beads, or microspheres are suspended in solution, so they are free to interact with the sample. This creates an assay platform that is - flexible (protein, peptides, DNA - reproducible ( as beads are uniformly distributed in solution) - improved sensitivity (better S/N, and near solution-phase kinetics) The object is to conjugate, or couple a different biological reaction to the surface of each bead set

20 Amine Coupling Kit

21 Amine Coupling Kit = o oH o + Chimica dell’accoppiamento C N H C N H
R H C o N R H + = Carbodiimide reaction Le beads vengono attivate dall’EDC (1-ethyl-3 (3-dimethylaminopropyl) carbodiimide hydrocloride) Reagiscono con S-NHS (N-hydroxysuccinimide) Si formano esteri stabili che reagiscono con il gruppo NH2 della proteina per formare un legame covalente

22 Amine Coupling Kit 171-406001 171-5060XX
Per 30 reazioni di accoppiamento XX COOH Beads Regione XX, 1 ml Per 10 reazioni di accoppiamento Un coupling kit e 3 ml di beads sono sufficienti per 60 multiwell da 96 pozzetti

23 Saggi di Interazione DNA - Proteine

24 Interazione DNA-Proteine