Necrosis related HIF-1α expression predicts prognosis in patients with endometrioid endometrial carcinoma

BMC Cancer

Table 2 Overview of the antibodies used and tissue processing details.

Primary Anti Body	Type Anti Body	Source	Dilution	Antigen Retrieval	Second step	Positive control	Incubation time/temp (primary antibody)	Procedure
HIF-1α	MoAb	Transduction	1:50	EDTA, Ph9.0, 20 minutes, 93°C	NV	mamma	o/n 4°C	By hand
Glut-1	PoAb, Rabbit	DAKO	1:200	Citrate, Ph6.0, 20 minutes, 93°C	G-AR IgG + strep 1	Placenta	60 minutes/room temp	Autostainer
CAIX	PoAb, Rabbit	Abcam	1:1000	Citrate, Ph6.0, 20 minutes, 93°C	PV	Grawitz tumour	60 minutes/room temp	By hand
p27^kip1	MoAb	Transduction	1:500	Citrate, Ph6.0, 20 minutes, 93°C	PV	Skin	o/n 4°C	By hand

HIF-1α = hypoxia-inducible factor-1α; Glut-1 = glucose transporter-1; CAIX = carbonic anhydrase IX; MoAb = monoclonal antibody; PoAb = polyclonal antibody; Transduction = BD Transduction Laboratories, BD Biosciences, San Diego, CA, USA; DAKO = DAKOCytomation, Glostrup, Denmark; Abcam = Abcam, Cambridge Science Par, Cambridge, UK; G-aR IgG = biotinylated Goat-anti Rabbit IgG (BA-1000, Vector laboratories, CA, diluted 1:500) + strep 1 = Streptavidin peroxidase labelling (Streptavidin HRP IM0309, Beckman Coulter, diluted 1:100); NV = Novolink™ polymer (Novocastra); PV = Powervision ready to use (Poly-HRP-anti Ms/Rb/RtlgG biotin free, ImmunoLogic, ImmunoVision technologies, Brisbane CA, USA).

ISSN: 1471-2407