Tissue was fixed in neutral 10% buffered formaldehyde, embedded in paraffin and cut into 5-μm-thick slices for ERO1-α staining and CD31 staining. Reactivity of the anti-ERO1-α mAb was determined by perinuclear staining within tumour cells, indicating endoplasmic reticulum localisation. Anti-CD31 polyclonal antibody, which was used for staining of blood vessels in tumour tissues obtained from mice, was purchased from Abcam (Cambridge, MA, USA).
Anti cd31 polyclonal antibody
Manufactured by Abcam
Sourced in United States, United Kingdom
The Anti-CD31 polyclonal antibody is a laboratory reagent used for the detection and analysis of the CD31 protein. CD31, also known as PECAM-1, is a cell adhesion molecule expressed on the surface of various cell types, including endothelial cells, platelets, and some leukocytes. This antibody can be used in techniques such as immunohistochemistry, immunocytochemistry, and flow cytometry to identify and study cells expressing CD31.
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4 protocols using anti cd31 polyclonal antibody
1
Immunohistochemical Staining of Tumor Tissues
2
Quantifying Muscle Angiogenesis and VEGF
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The major thighs and calf muscles (including the gracilis, abductors, adductors, gastrocnemius, and soleus) were excised immediately following each imaging time point and were fixed in neutral buffered formalin. Sections were taken throughout the muscle samples at 12 μm thickness and stained for CD31 using anti-CD31 polyclonal antibody (Abcam Singapore Pte., Ltd.) or for VEGF using anti-VEGF monoclonal antibody (Abcam Singapore Pte., Ltd.). Muscle capillary number was calculated as CD31-positive vessel number and VEGF levels as VEGF-positive cells per 10 fields of view assessed per sample. Binding was analysed manually as previously described [7 (link)] in a manner blinded to treatment assignment.
3
Evaluation of Tumor Responses to 90Y-1849 Therapy
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As a separate experiment, tumor samples (n = 3/time-point) were extirpated at days 1, 2, and 5 after intravenous injection of intact 1849 (0 MBq) or 3.7 MBq of 90Y-1849 with saline or SQAP (2 mg/kg body weight). Tumor sections were stained with hematoxylin and eosin (Sakura Finetek USA, Torrance, CA, USA). Apoptotic cells in tumors were stained by TUNEL staining with a DeadEnd Colorimetric TUNEL system (Promega, Madison, WI, USA). Ki-67 antigen was detected using an anti-human Ki-67 polyclonal antibody (Agilent Technologies Japan, Tokyo, Japan) as described previously [38] (link). CD31 antigen was detected using an anti-CD31 polyclonal antibody (Abcam, Cambridge, UK) as described previously [23] . Details are provided in the Supplementary information.
4
Retinal Vascular Staining and Inflammation
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