HTRF assays for Oncology and Inflammation SIGNALING IN THE IMMUNE SYSTEM NK / NKT CELLS Cytokines Markers GM-CSF (CSF2) IL10 Granzyme B IFNg TNFa Perforin-1 Pi3 Syk AT ME SLP6 Iβ ER C-Raf NFkB Cisbio cell signaling assays available IMMUNE CHECKPOINTS Biochemical assays to identify checkpoint blockers TCR MHC II PD-1 PD-L1 CT C C CT C C G C MC II C SIα A2AR enosine T CELL SIGNALING TCR CD 4/8 CTLA4 PD1 CD3 CD28 CD40L AP0 SHP1 PLC SLP6 p38 c-Raf AT IP1 ME GS3β NFAT Iβ ER NFB AP1 nhiitory INγ I I I I TNα timulatory (Cisio cytokine assays availale) DNA Cisio cell signaling assays availale pSLP6 Dexamethasone inhibitory effect on CD3/CD28 stimulated PBMCs 8000 10000 ** hIFNγ L m 6000 / g hIL2 p ] s 5000 4000 e n 2000 i k o t HTRF signal 2000 y C 1000 Neg [ 0 0 Basal 4 40 CD3/CD28 anti-CD3 Ab (g/mL) Dexamethasone Pathway monitoring in primary T cells upon CD3 stimulation Cytokine panel on PBMCs (300,000 cells per well, 5 min CD3 stimulation) B CELL SIGNALING BCR PD1 CD28 CTLA4 CD40L BT SHP1 AT PLC BLN Iβ GS3β c-Raf IP1 ME NFAT NFB ER nhiitory AP1 INγ I I I I I TNα timulatory (Cisio cytokine assays availale) DNA Cisio cell signaling assays availale Stimulation effect of pervanadate on Raji cells Dexamethasone inhibitory effect on LPS stimlated PBMCs 15000 S/B = 10.7 40000 25000 o H h 20000 T hIL6 L p R s 30000 m / F o 15000 10000 g hIL1β h R p P a ] t 10000 s o i i 20000 hTNFα o t e 4000 a n T i o R k 5000 t o F a 10000 t y R l 2000 T C [ H 0 0 0 0 15 30 60 0 15 30 60 LPS [Pervanadate] (µM) Dexamethasone Pervanadate effect on BTK accumulation (phospho & total) Cytokine panel on PBMCs INNATE IMMUNITY Cytokines Inflammation markers IL1b CCL3 (MIP1a) Histamine IL6 CCL4 (MIP1b) PGE2 IL8 CCL5 (RANTES) LTC4 IFNb TNFa LTB4 CXCL10 (IP-10) TLR5 TLR10 TLR2/TLR6 TLR1/TLR2 TLR4 dsDNA cGAS MYD88 ATP + GTP 2’-3’ cGAMP TBK1 IRAK3 IRAK1 IRAK2 STING IRAK4 IRF3 STING p38 IKKβ TAK1 JNK NFκB NFκB IRF3 IRF3 TNFα, IL6, IL1β Type-I-IFNs Cisio cell signaling assays availale Toll-like receptor STING signaling pathway signaling pathway CELLULAR PHOSPHORYLATION ASSAYS Transfer ncuate supernatant Plate timulate T detection 3 yse antiodies • Cell-based assays • Compatible with cell lysates, tissue extracts, iPS cells, 3D cell models • Sample volume 16 μL • More sensitive than Western Blot • < 1 hr bench time • Pharmacological studies of pathway inhibitors, agonists, and allosteric modulators CELL-BASED CYTOKINE ASSAYS ncuate sample T detection antiodies • Sample volume 16 μL • < 2 hrs bench time • Validated on PBMCs and cell lines (THP1, Jurkat...) • Compatible with Cisbio 96-well low-volume plate • Allow multiple cytokine measurements from 1 sample MDB008 - 01/2020 - Copyright Cisbio Bioassays. All rights reserved. HTRF and Tag-lite are registered trademarks of Cisbio Bioassays. Europe and other countries +33(0)466 796 705 | [email protected] U.S.A. and Canada 1-888-963-4567 | [email protected] China +86 21 5018 9880 | [email protected] Japan +81-(0)43-306-8712 | [email protected] www.cisbio.com.