Immunohistochemical Analysis of NPNT and CYP11B2

Immunohistochemistry was carried out using the peroxidase-antiperoxidase method on fresh frozen human tissue. In cases where fresh frozen tissue was unavailable, immunohistochemistry was performed on formalin-fixed, paraffin-embedded adrenal sections (4 μm) using an automated immunostainer with cover tile technology (Bond-III system, Leica Biosystems). NPNT antibody (HPA003711, Sigma; 1:50 dilution), and CYP11B2 antibody (custom mouse anti-human antibody from Dr Celso E. Gomez-Sanchez)22 (link) were used as the primary antibodies. Further details are provided in the Data Supplement.

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Teo A.E., Garg S., Johnson T.I., Zhao W., Zhou J., Gomez-Sanchez C.E., Gurnell M, & Brown M.J. (2017). Physiological and Pathological Roles in Human Adrenal of the Glomeruli-Defining Matrix Protein NPNT (Nephronectin). Hypertension (Dallas, Tex. : 1979), 69(6), 1207-1216.

Publication 2017

Bond-III system HPA003711

Anti antibody Antibodies Antibody Dilution Embedded paraffin Formalin Frozen Human Immunohistochemistry Mouse Peroxidase Supplement Tissue

Corresponding Organization :

Other organizations : Hutchison/MRC Research Centre, Addenbrooke's Hospital, University of Cambridge, Wellcome Trust, MRC Cancer Unit

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Variable analysis

independent variables

Tissue type (fresh frozen vs. formalin-fixed, paraffin-embedded)
Primary antibody type (NPNT antibody and CYP11B2 antibody)

dependent variables

Immunohistochemistry staining patterns

control variables

Tissue thickness (4 μm for formalin-fixed, paraffin-embedded sections)
Immunohistochemistry method (peroxidase-antiperoxidase method for fresh frozen tissue, automated immunostainer with cover tile technology for formalin-fixed, paraffin-embedded tissue)

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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