Sections from cultured islets and islets + DPS preparations were double stained by immunofluorescence for apoptotic nuclei with the terminal deoxynucleotidyl transferase biotin-dUTP nick-end labeling (TUNEL) technique (ApopTag Plus Fluorescein In Situ Apoptosis Detection Kit; Merk Millipore), and for insulin using a rabbit anti-human insulin antibody (Santa Cruz Biotechnology Inc., Dallas, TX, USA) (final dilution 1:100). Preparations were visualized by immunofluorescence with donkey anti-rabbit IgG conjugated with Alexa Fluor 555 (Life Technologies) (final dilution 1:400). Nuclei were stained with 300 nM DAPI (Life Technologies). β-cell apoptosis was expressed as percentage of TUNEL-positive β-cells with pyknotic nucleus. A minimum of 1,200 cells per sample was counted.
Rabbit anti human insulin antibody
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Rabbit anti-human insulin antibody is a laboratory reagent used to detect and quantify human insulin in various biological samples. It is a polyclonal antibody produced by immunizing rabbits with human insulin. This antibody can be utilized in techniques such as Western blotting, ELISA, and immunohistochemistry to identify and measure insulin levels.
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2 protocols using rabbit anti human insulin antibody
1
Quantifying Pancreatic β-Cell Apoptosis
2
Quantifying Pancreatic Cell Proliferation
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Isolated pancreata were immersion-fixed in 4% paraformaldehyde. Tissue was then routinely processed for paraffin embedding, and 5-µm sections mounted on glass slides were immunostained with rabbit anti-human insulin antibody (diluted 1:1000; Santa Cruz Biotechnology Inc., Santa Cruz, CA). BrdU incorporation was analysed as described previously [25] . The sections were double-immunostained with anti-BrdU antibody (diluted 1:10; BD Biosciences, Franklin Lakes, NJ, USA) and anti-insulin antibody (diluted 1:1000). BrdU-positive beta cells were quantitatively assessed as a percentage of the total number of beta cells and at least 50 islets per mouse were counted in each group.
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