Immunohistochemical Analysis of Angiogenic Factors

Streptavidin alkaline phosphatase (APase) immunohistochemistry was performed on cryopreserved tissue sections using a nitroblue tetrazolium (NBT) development system as previously described [21 (link)]. An ABC system (Dako) was performed in paraffin-embedded mouse tissue as previously described [21 (link)]. Primary antibodies used include: HIF-1α (ABCAM), ANGPTL-4 (ABCAM), VEGF (Santa Cruz) after dilution in TBS with 1% bovine serum albumen (BSA). All immunohistochemical reagents, including antibodies, were identical for all specimens.

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Hu K., Babapoor-Farrokhran S., Rodrigues M., Deshpande M., Puchner B., Kashiwabuchi F., Hassan S.J., Asnaghi L., Handa J.T., Merbs S., Eberhart C.G., Semenza G.L., Montaner S, & Sodhi A. (2016). Hypoxia-inducible factor 1 upregulation of both VEGF and ANGPTL4 is required to promote the angiogenic phenotype in uveal melanoma. Oncotarget, 7(7), 7816-7828.

Publication 2016

ABC system ANGPTL-4 VEGF

Alkaline phosphatase Antibodies Bovine serum albumen Dilution Embedded paraffin Immunohistochemistry Mouse Nitroblue tetrazolium Streptavidin Tissue Vegf

Corresponding Organization : Johns Hopkins Medicine

Other organizations : First Affiliated Hospital of Chongqing Medical University, Chongqing Medical University, University of Maryland, Baltimore

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Variable analysis

independent variables

Primary antibodies used: HIF-1α (ABCAM), ANGPTL-4 (ABCAM), VEGF (Santa Cruz)

dependent variables

Immunohistochemical analysis of cryopreserved tissue sections using a nitroblue tetrazolium (NBT) development system
Immunohistochemical analysis of paraffin-embedded mouse tissue using an ABC system (Dako)

control variables

All immunohistochemical reagents, including antibodies, were identical for all specimens.

controls

Positive control: None specified.
Negative control: None specified.

Annotations

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