Pancreata were fixed in 4% paraformaldehyde for 72 hr, dehydrated in sucrose and mounted in OCT. Ten‐micron sections were cut on a Leica cryostat onto Polysine slides (VWR), air‐dried and fixed again for 10 min in acetone. Guinea‐pig‐anti‐insulin antibody (DAKO) was detected with anti‐guinea‐pig Alexa 546 (Molecular Probes, Eugene, OR). Anti‐CD4 (BD) was detected with anti‐rat Alexa 488 (Molecular Probes). GLUT‐1 in cells was detected using ab15309 (Abcam, Cambridge, UK) and Alexa 647 anti‐rabbit (Molecular Probes) after permeabilization with PBS‐Triton‐X. Nuclei were visualized with DAPI (Molecular Probes). The sections were viewed with a confocal microscope (Zeiss, Oberkochen, Germany) and processed using zen software.
Anti guinea pig alexa 546
Manufactured by Thermo Fisher Scientific
Anti‐guinea‐pig Alexa 546 is a fluorescently labeled secondary antibody that specifically binds to guinea pig primary antibodies. The Alexa 546 fluorophore is attached to the secondary antibody, allowing for the detection and visualization of guinea pig antigens in various applications such as immunofluorescence and flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions)
Polysine slides, by Avantor (2 mentions)
Guinea pig anti insulin antibody, by Agilent Technologies (2 mentions)
Anti cd4, by BD (1 mentions)
Ab15309, by Abcam (1 mentions)
Anti mouse cd4, by BD (1 mentions)
Lsm 700 confocal, by Zeiss (1 mentions)
Cryostat, by Avantor (1 mentions)
2 protocols using anti guinea pig alexa 546
1
Pancreatic Cryosectioning and Immunofluorescence
2
Immunohistochemical Analysis of Pancreatic Islets
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Pancreata and graft-bearing pinnae were fixed in 4% PFA for 72 h, dehydrated in sucrose, and mounted in OCT. 10 µm sections were cut on a Leica cryostat and placed on Polysine slides (VWR), air dried, and fixed again for 10 min in acetone. Sections were stained using guinea pig-anti-insulin antibody (DAKO) at a 1:40 dilution and detected with anti-guinea pig Alexa 546 (Molecular Probes). Rabbit anti-human glucagon (Millipore) was used at a 1:50 dilution and detected with anti-rabbit Alexa 488 (Molecular Probes). Anti-mouse CD4 (BD) was detected with anti-rat Alexa 488 (Molecular Probes). Nuclei were visualized with DAPI (Molecular Probes). The sections were viewed using either a Zeiss Axioskop 2 or LSM700 Confocal (Zeiss) and processed using Zen software.
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