PARP enzymatic assay
⦁ PARP1& PARP2 & PARP3 enzymatic activity assay
⦁ PARP7 enzymatic activity assay
⦁ PARP5a/TNKS1,PARP5b/TNKS2, PARP10, PARP14, PARP15 enzymatic activity assay
⦁ PARP trapping assays on PARP1 and PARP2
High Content Imaging assay
⦁ PARylation inhibition on isogenic A549 cells(in A549 wild type (WT),
PARP1-KO and PARP2-KO cells)
⦁ PARylation inhibition on MDA MB-436 (BRCA1 mutant) cancer line
⦁ PARP-1 trapping by using PARP-1 and gH2A.X antibody pair on MDA-MB-436
(BRCA1 mutant) cancer line
⦁ gH2AX% induction determination by treating MDA-MB-436 (BRCA1 mutant) cancer line
with 10uM MMS
⦁ gH2AX% induction determination by treating DLD1 WT and DLD1 BRCA2-/- cell lines
detected by HCI
⦁ PARPi induces an accumulation of cytosolic DNA and activates STING signaling pathway
by HCI in MDA-MD-436
Pharmacodynamic (PD) assay
⦁ Anti-tumor efficacy of AZD5305 in BRCA1 mutation tumor model
→ Customized in-house PARyation Elisa assay
→ Human Phospho-Histone H2AX (S139) Elisa assay
⦁ Anti-tumor efficacy of RBN2397 in NCI H1373 tumor model
→ Human IFN-b Elisa assay
→ Human CXCL-10 Elisa assay
→ We also can measure SQSTM1(p62) and IL-b levels.
7 day cell viability CTG assay
⦁ MDA-MB-231, MDA-MB 436, DLD1, BRCA2-/-DLD1
⦁ NCI-H1373 and CT-26 cell line
Colony formation CFU assay
⦁ MDA-MB-231, MDA-MB-436, DLD1, BRCA2-/-DLD1
Others
⦁ gH2AX% induction determination by treating MDA-MB-436 (BRCA1 mutant) cancer line
with 10uM MMS detected by FACS
⦁ Regulation on p STAT1 by Immunoblotting
⦁ Regulation on IFN-b by Elisa assay in below cell lines : NCI-H1373, CT26 and MC38, etc..
⦁ Regulation on CXCL-10 by Elisa assay in below cell lines : NCI-H1373, CT26 and MC38, etc..
⦁ Regulation on CXCL-10 by RT-PCR in NCI-H1373 cell line. |