MCF-7 CDX |
Ki67 (MIB-1 clone, #M7240, Dako) 1:100, 60 mins, RT |
pH6 antigen retrieval solution (Dako), 110 oC at pressure for 2 mins |
Envision+ anti-mouse secondary (Dako), 30 mins, RT |
Cleaved CK18 (M30 Cytodeath, #12140322001, Roche) 1:200, 60 mins, RT |
pH6 antigen retrieval solution (Dako), 110 oC at pressure for 5 mins |
Envision+ anti-mouse secondary (Dako), 30 mins, RT |
ERα (ID5 clone, #M704701, Dako) 1:200, 60 mins, RT |
pH6 antigen retrieval solution (Dako), 110 oC at pressure for 2 mins |
Biotinylated anti-mouse secondary (Dako) 30 mins, RT ABC amplification step (Vector Labs) 30 mins, RT |
HIF1α (#610959, BD Biosciences) 1:50, 120 mins, RT |
pH8 EDTA solution, 110 oC at pressure for 2 mins |
Envision+ anti-mouse secondary (Dako), 30 mins, RT |
E-cadherin (24E10 clone, #3195, Cell Signaling Technology) 1:100, 60 mins, RT |
pH6 antigen retrieval solution (Dako), 110 oC at pressure for 2 mins |
Biotinylated anti-rabbit secondary (Dako) 30 mins, RT ABC amplification step (Vector Labs) 30 mins, RT |
phospho-γH2AX (#2577, Cell Signaling Technology) |
pH9 retrieval solution (Dako), 110 oC at pressure for 2 mins |
Envision+ anti-rabbit secondary (Dako), 30 mins, RT |
F4/80 (Serotec, #MCA497G) 1:200, 60 mins, RT |
pH6 antigen retrieval solution (Dako), 110 oC at pressure for 2 mins |
Biotinylated anti-rat secondary (Dako) 30 mins, RT ABC amplification step (Vector Labs) 30 mins, RT |
LungPDX and CDX |
Ki67 (MIB-1 clone, #M7240, Dako) 1:75, 30 mins, RT |
pH6 antigen retrieval solution (Dako), steam heat for 30 mins |
pH6 antigen retrieval solution (Dako), steam heat for 30 mins |
Cleaved Caspase 3 (#9661, Cell Signaling) 1:75, 30 mins, RT |
pH6 antigen retrieval solution (Dako), steam heat for 30 mins |
Envision+ anti-mouse secondary (Dako), 30 mins, RT |
Prostate PDX |
Ki67 (MIB-1 clone, #M7240 Dako) 1:100, over night, 4°C |
10 mM Na-citrate, pH6, at 99 oC for 20 |
DAKO EnVision Detection Kit, Peroxidase/DAB, Rabbit/Mouse; 30 mins. RT, DAB 2 × 5 mins |
AR (#sp107, Cell Marque) 1:200, over night, 4°C |
10 mM Na-citrate, pH6, at 99 oC for 20 |
DAKO EnVision Detection Kit, Peroxidase/DAB, Rabbit/Mouse; 30 mins. RT, DAB 2 × 5 mins |