In order to verify whether the SDF-1α has been coated on the scaffold, immunofluorescent staining was performed by using SDF-1α antibody (Boster, China; diluted 1:50) and incubated overnight at 4 °C in a humidified chamber. The secondary antibody was applied with an Alexa Fluor 488-labled donkey anti-rabbit IgG (Molecular Probes, USA; diluted 1:500). Three samples for each group were imaged by the fluorescence microscope (Olympus IX71, Japan).
Alexa fluor 488 labled donkey anti rabbit igg
Manufactured by Thermo Fisher Scientific
Sourced in United States
Alexa Fluor 488-labeled donkey anti-rabbit IgG is a secondary antibody conjugate designed for use in immunodetection and immunolabeling techniques. It is specific for rabbit immunoglobulin G (IgG) and is labeled with the Alexa Fluor 488 fluorescent dye, which has an excitation maximum of 495 nm and an emission maximum of 519 nm.
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2 protocols using alexa fluor 488 labled donkey anti rabbit igg
1
Immunofluorescent Staining of SDF-1α on Scaffold
2
Immunofluorescence Analysis of Stem Cell Markers
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After subcutaneous implantation for 5 days, 10 days, 28 days and 56 days, samples were fixed in 4% paraformaldehyde, and then decalcified in 12% EDTA-2Na. The sample was cut in half, and one for paraffin sections while the other for frozen sections. Frozen sections (8 μm thick) were then subjected to immunofluorescence for antibody of rabbit anti-rat CD29, CD34, CD44, CD68, CD90, and CD105 (Bioss, China; diluted 1:100). The Alexa Fluor 488-labled donkey anti-rabbit IgG (Molecular Probes, USA; diluted 1:500) was used as the second antibody. DAPI (1 μg/mL) staining was used to reveal the nuclei.
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