Naïve or injected embryos were fixed and labeled with monoclonal anti–α-Tubulin (DM1A; Sigma-Aldrich) followed by the secondary antibody Alexa Flour 488 anti-mouse (A21202, Invitrogen) and Hoechst (H-3570 Invitrogen). Embryos were dissected to remove the yolk and mounted in fluoromount-G (SouthernBiotech) on a coverslip with the animal pole facing the coverslip. Samples were imaged by confocal microscopy as described below. Detailed protocols are provided in SI Appendix, Supplementary Methods .
Alexa flour 488 anti mouse
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
Alexa Fluor 488 anti-mouse is a fluorescently labeled secondary antibody used to detect and visualize mouse primary antibodies in various biological applications, such as immunofluorescence, flow cytometry, and Western blotting. It provides bright, specific labeling with minimal background signal.
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2 protocols using alexa flour 488 anti mouse
1
Embryo Immunofluorescence Labeling Protocol
2
Immunofluorescent Analysis of Pancreatic Islets
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Paraffin-embedded sections (5-µm-thick) of the pancreatic splenic lobe were subjected to fluorescence immunohistochemistry as described previously (28 (link)) using a 1:200 dilution of mouse monoclonal anti-insulin antibody (I2018, Sigma Aldrich, St. Louis, MO, USA) or rabbit polyclonal antibodies against the proliferation marker, Ki67 (ab66155, Abcam, UK), and a 1:500 dilution of Alexa Flour 488 anti-mouse or Alexa Flour 555 anti-rabbit secondary antibodies (Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA). Sections were cover-slipped with Vectashield mounting medium containing DAPI (Vector Laboratories, Inc., Burlingame, CA, USA) and observed under fluorescence microscopy (Microscope BX60F5, Olympus) using the 40X objective, and the fluorescence signal was quantified from at least 15 islets per animal using the Image-Pro Plus 5.1 software.
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