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

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

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
p27kip1 MoAb Transduction 1:500 Citrate, Ph6.0, 20 minutes, 93°C PV Skin o/n 4°C By hand
  1. 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).