Table 2.

Details of the antibodies, dilutions, and antigen retrieval methods used in this study

Primary antibody	Specificity	Antigen retrieval	Dilution/incubation time	Source	Reference
ESEE 122	HIF-1α	0.1 m EDTA 3 min, PC	Neat 90 min, RT	In house	27
EP 190b	HIF-2α	0.1 m EDTA 60°C, ON	Neat 90 min, RT	In house	27
CW27	DEC-1	Citrate buffer 10 min, MW	1:1000 45 min, RT	Prof. Adrian Harris, CRUK	18
VG-1	VEGF	0.05 m Tris/0.2 mm EDTA 3 min, PC	1:4, 90 min, RT	In house	28
C-17	VEGF-R1	As above	1:200 90 min, RT	Santa Cruz, CA
Y-23	VEGF-R2	As above	1:200 90 min, RT	Santa Cruz, CA
pKDR34	pKDR	Citrate buffer 3 min, MW	1:5 90 min, RT	In house	29

All primary antibodies were visualized with the Envision-HRP/AP kits (Dako, Ely, UK) with the exception of ESEE 122, which was visualised with the TSA-HRP kit (NEN, Boston, MA, USA).

EDTA, ethylenediamine tetraaceticacid; ON, overnight; RT, room temperature; PC, pressure cooker; MW, microwave; CRUK, Cancer Research, UK.