Immunohistochemical analysis was conducted on formalin‐fixed paraffin‐embedded tissues. Colon sections were stained with antibodies specific for β‐catenin (Cell Signaling Technology), BrdU (AbD Serotec), cleaved caspase‐3 (Cell Signaling Technology), CD31 (Dianova), c‐MYC (Santa Cruz Biotechnology), cyclin D1 (Santa Cruz Biotechnology), F4/80 (Life Technologies), iNOS (Abcam), and Ki‐67 (Dako Deutschland GmbH). The antibody‐antigen complexes were detected using biotinylated donkey anti‐rat and donkey anti‐rabbit secondary antibodies (Dianova) and the Dako REAL Detection System (Dako). Immunohistochemical detection of HIF‐1α (Cayman Chemical) was performed as previously described.23 Nuclei were counterstained with hematoxylin. Negative controls were performed by omitting the primary antibody. The average number of positively stained cells within at least six high power fields (HPF, 0.237 mm2) was determined by a blinded independent investigator.
Biotinylated donkey anti rat and donkey anti rabbit secondary antibodies
Manufactured by Dianova
Sourced in Germany
Biotinylated donkey anti-rat and donkey anti-rabbit secondary antibodies are laboratory reagents used in various immunoassay techniques. These antibodies are specifically designed to detect and bind to primary antibodies raised in rat or rabbit, respectively. The biotin label allows for further detection and signal amplification in the assay. The core function of these secondary antibodies is to facilitate the identification and quantification of target proteins or molecules in a sample.
Automatically generated - may contain errors
Lab products found in correlation
Dako real detection system, by Agilent Technologies (2 mentions)
Hif 1α, by Cayman Chemical (2 mentions)
Bromodeoxyuridine (brdu), by Bio-Rad (1 mentions)
F4 80, by Thermo Fisher Scientific (1 mentions)
C myc, by Santa Cruz Biotechnology (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
β catenin, by Cell Signaling Technology (1 mentions)
Cyclin d1, by Santa Cruz Biotechnology (1 mentions)
F4 80, by Bio-Rad (1 mentions)
Fsp 1, by Agilent Technologies (1 mentions)
α sma, by Merck Group (1 mentions)
2 protocols using biotinylated donkey anti rat and donkey anti rabbit secondary antibodies
1
Immunohistochemical Analysis of Colon Tissues
2
Immunohistochemical Analysis of Intestinal Inflammation
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Immunohistochemical analysis was conducted on formalin-fixed paraffin-embedded tissues.
For histopathological evaluation, intestinal sections were stained with hematoxylin and eosin and mucosal inflammation was graded as described (66) . Intestinal sections were also stained with antibodies specific for F4/80 (Bio-Rad, Hemel Hempstead, UK), FSP-1 (Dako, Hamburg, Germany), HIF-1α (Cayman Chemical, Ann Arbor, USA), αSMA (Merck Millipore, Darmstadt, Germany) and ZO-1 (Thermo Fisher Scientific, Waltham, USA). The antibodyantigen complexes were detected using biotinylated donkey anti-rat and donkey anti-rabbit secondary antibodies (Dianova, Hamburg, Germany) and the Dako REAL Detection System (Dako). Sections were counterstained with hematoxylin. Immunohistochemical detection of HIF-1α was performed as previously described (77) . For the immunohistochemical assessment of hypoxic areas, the Hypoxyprobe-1 Kit (Hypoxyprobe Inc., Burlington, USA) was used as described before (78) . Tumor-associated macrophage abundance was determined by quantification of F4/80+ cells in at least 5 random fields of view.
For histopathological evaluation, intestinal sections were stained with hematoxylin and eosin and mucosal inflammation was graded as described (66) . Intestinal sections were also stained with antibodies specific for F4/80 (Bio-Rad, Hemel Hempstead, UK), FSP-1 (Dako, Hamburg, Germany), HIF-1α (Cayman Chemical, Ann Arbor, USA), αSMA (Merck Millipore, Darmstadt, Germany) and ZO-1 (Thermo Fisher Scientific, Waltham, USA). The antibodyantigen complexes were detected using biotinylated donkey anti-rat and donkey anti-rabbit secondary antibodies (Dianova, Hamburg, Germany) and the Dako REAL Detection System (Dako). Sections were counterstained with hematoxylin. Immunohistochemical detection of HIF-1α was performed as previously described (77) . For the immunohistochemical assessment of hypoxic areas, the Hypoxyprobe-1 Kit (Hypoxyprobe Inc., Burlington, USA) was used as described before (78) . Tumor-associated macrophage abundance was determined by quantification of F4/80+ cells in at least 5 random fields of view.
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