Formalin-fixed paraffin-embedded xenografts were cut into 6 μm thick sections, and immunochemical analysis was conducted using the Vectastain Elite ABC Universal kit (Vector Laboratories, Burlingame, CA, USA) using a previously described methodology [14 (link),22 (link)]. Antibodies against CD31 (1:20, Dianova, Geneva, Switzerland), lymphatic vessel hyaluronic acid receptor-1 (LYVE-1) (1:100; DAKO, Glostrup, Denmark), and Ki-67 nuclear antigen (1:100; Dako, Glostrup, Denmark) were used for the analysis. For each marker, whole-surface staining was quantified using Image J software (NIH, Bethesda, MD, USA).
Ki 67 nuclear antigen
Manufactured by Agilent Technologies
Sourced in Denmark
The Ki-67 nuclear antigen is a protein that is expressed during all active phases of the cell cycle, with the exception of the resting phase (G0). It is commonly used as a marker for cell proliferation and is widely employed in the field of pathology to assess the growth fraction of a given cell population.
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3 protocols using ki 67 nuclear antigen
1
Immunochemical Analysis of Xenograft Tissues
2
Quantifying Tumor Angiogenesis and Proliferation
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Sections (6 μm) were cut from formalin-fixed paraffin-embedded xenografts. Immunohistochemistry was performed using the Vectastain Elite ABC Universal kit (Vector Laboratories, Burlingame, CA) as described previously [23 (link), 64 (link)]. Antibodies against CD31 (1:20, Dianova), Ki-67 nuclear antigen (1:100; Dako), and cleaved caspase-3 (1:100; BD Pharmingen™) were used for the analysis. For each marker, whole-surface staining was quantified using Image J Software (NIH, Bethesda, USA).
3
Immunohistochemical Analysis of MSTO-211H Xenografts
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Sections (6 mm thick) were cut from formalin-fixed paraffin-embedded MSTO-211H xenografts. Immunohistochemistry was performed using the Vectastatin Elite ABC Universal kit (Vector Laboratories, Burlingame, CA) as described previously. 20, 28 Antibodies recognizing CD31 (1:20, Dianova), Ki67 nuclear antigen (1:100; Dako), cleaved caspase-3 (1:100; BD Pharmingen), and lymphatic vessel hyaluronic acid receptor-1 (LYVE-1) (1:100) were used for our analysis. For each marker, whole-surface staining was quantified using CALOPIX Software (TRIBVIN Medical, Châtillon, France).
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