Astrocytes were fixed in 4% paraformaldehyde and permeabilized with 0.2% Triton X-100. Following overnight incubation with Ki67 antibodies (Abcam, Cambridge, MA), visualization was performed following incubation with Alexa Fluor 488 IgG secondary antibodies (Invitrogen, Carlsbad, CA). Cells were counterstained with DAPI. For each independent culture, at least five distinct microscopic fields were analyzed on a Nikon Eclipse TE300 fluorescence inverted microscope (Nikon, Tokyo, Japan) equipped with an optical camera (Optronics, Goleta, CA) and MetaMorph image analysis software (Molecular Devices, Dowingtown, PA).
Alexa fluor 488 igg secondary antibodies
Manufactured by Thermo Fisher Scientific
Sourced in United States
Alexa Fluor 488 IgG secondary antibodies are fluorescent-labeled antibodies designed to detect and visualize target proteins in various immunoassays and microscopy applications. These antibodies are conjugated with the Alexa Fluor 488 dye, which exhibits bright green fluorescence when excited by a suitable light source.
Automatically generated - may contain errors
2 protocols using alexa fluor 488 igg secondary antibodies
1
Astrocyte Proliferation Assay via Ki67
2
Apoptosis Markers in Epithelial and Fibroblast Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Primary gastric epithelial cells and guinea pig fibroblasts after exposure to blank MPs: CHI, CHI-Plur or CHI-GlcNAc at the concentration of 10 mg/mL or 5 mg/mL, or E. coli LPS (as a positive control), were immunohistochemically stained for the presence of early pro-apoptotic caspase 3 (CC3) or middle apoptosis stage caspase 9 (CC9) as well as late apoptosis protein carbamoyl-phosphate synthetase 2 aspartate transcarbamylase and dihydroorotase (CAD). The procedure of staining was performed using fluorescently labelled rabbit-specific primary antibodies (Santa Cruz Biotechnology, Dallas, TX, USA), and then anti-rabbit Alexa Fluor 488-IgG secondary antibodies (Invitrogen, Waltham, MA, USA), as previously described31 (link). The amount of CC9 and CAD was determined by measuring fluorescence intensity at 495 nm excitation and 519 nm emission, using a SpectraMaxi3 reader (Molecular Devices, San Jose, CA, USA). Four independent experiments were carried out in triplicate for each experimental variant.
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