Immunohistochemical analysis of placental HIF-1α

Sections (4 μm) of paraffin-embedded placental tissue were stained as described [52 (link)] with antigen retrieval at pH6.0 for 20 min. Sections were incubated with antibodies against HIF-1α (Abcam, Cambridge, UK) at 2 μg/mL over night at 4 °C. IHC staining was detected using the ENVISION™ system (DAKO, Santa Clara, CA, USA) with NovaRED™ (Vector Labs, Burlingame, CA, USA) as the substrate. Tissue sections were counterstained with Harris haematoxylin. Images were captured using NIS Element software (Nikon, Tokyo, Japan) via Nikon Digital Sight (H550S, Nikon, Tokyo, Japan).

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McCracken S.A., Seeho S.K., Carrodus T., Park J.H., Woodland N., Gallery E.D., Morris J.M, & Ashton A.W. (2022). Dysregulation of Oxygen Sensing/Response Pathways in Pregnancies Complicated by Idiopathic Intrauterine Growth Restriction and Early-Onset Preeclampsia. International Journal of Molecular Sciences, 23(5), 2772.

Publication 2022

antibodies against HIF-1α ENVISION™ system NovaRED™ NIS Element software

Antibodies Antigen Haematoxylin Paraffin Placental Sight Tissue Vector

Corresponding Organization : University of Sydney

Other organizations : Royal North Shore Hospital, University of Technology Sydney

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Variable analysis

independent variables

Antigen retrieval at pH6.0 for 20 min

dependent variables

HIF-1α expression

control variables

Paraffin-embedded placental tissue sections
Incubation with HIF-1α antibody at 2 μg/mL overnight at 4 °C
IHC staining using ENVISION™ system with NovaRED™ as substrate
Counterstaining with Harris haematoxylin

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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