For cell staining, cells grown on coverslips were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100 in TBS (50 mM Tris–HCl, pH 7.4, 150 mM NaCl) buffer. For specimen staining, formalin-fixed, paraffin-embedded sections of pancreatic cancer tissues were obtained. After blocking with 5% BSA (Yeasen, Shanghai) in PBS, antibodies against O-GlcNAc (Abcam, Inc., ab2739) or CA19-9 (Abcam, Inc., ab3982) at a dilution of 1/200 in 5% BSA were used to detect their expression at 4 °C overnight. After washing, the samples were stained with AlexaFluor® 488 goat anti-rabbit secondary antibody (Jackson ImmunoResearch, Inc.) or AlexaFluor® 594 goat anti-mouse secondary antibody (Jackson ImmunoResearch, Inc.) at a 1/200 dilution for 1 h at room temperature. DAPI (Southern Biotech, Inc.) was used as the nuclear counterstain. Fluorescent images were acquired using a Leica confocal laser scanning microscope (Leica, Inc.). Confocal imaging was performed in seven random fields. Mean fluorescence intensity (MFI) was calculated using Image-Pro plus software (version 6.0, Media Cybernetics, Inc.).
Alexafluor 488 goat anti rabbit secondary antibody
Manufactured by Jackson ImmunoResearch
Sourced in United States
AlexaFluor® 488 goat anti-rabbit secondary antibody is a fluorescent-labeled secondary antibody. It is designed to bind to rabbit primary antibodies, enabling their detection and visualization in various immunological assays.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 880, by Zeiss (2 mentions)
Ab2739, by Abcam (1 mentions)
Ab3982, by Abcam (1 mentions)
Confocal laser scanning microscope, by Leica (1 mentions)
Alexa fluor 594 goat anti mouse secondary antibody, by Jackson ImmunoResearch (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Southern Biotech (1 mentions)
Image pro plus software, by Media Cybernetics (1 mentions)
2 protocols using alexafluor 488 goat anti rabbit secondary antibody
1
Quantitative Immunofluorescence Staining of Pancreatic Cancer
2
Immunofluorescence Localization of Vesicle Proteins
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As described previously, LG samples were fixed in 4% PFA overnight at 4°C. The tissue blocks were washed, dehydrated, embedded in paraffin, and cut in 5 µm sections. Antigen retrieval was performed by incubation with a 0.01M citrate solution (pH 6.0) for 30 minutes at 98°C. The sections were then blocked with 0.3% BSA and followed by incubation with primary antibodies (Rabbit anti-RAB3d 1:200, Rabbit anti-Vamp8 1:100, and Rabbit anti-Snap23 1:100) and then with Alexa-Fluor 488 goat anti-rabbit secondary antibody (1:1000). Staining was visualized with a confocal Zeiss microscope (LSM-880, Zeiss, Oberkochen, Germany). High-resolution digital images were captured and stored in TIFF format. Antibodies used are rabbit anti-Rab3d polyclonal antibody (Catalog No. 12320-1-AP; Proteintech, Rosemont, IL, USA); rabbit anti-Vamp8 monoclonal antibody (Catalog No. MA5-32502; Invitrogen, Grand Island, NY, USA); rabbit anti-Snap23 polyclonal antibody (Catalog No. DF13314; Affinity Biosciences, Cincinnati, OH, USA); and Alexa-Fluor 488 goat anti-rabbit secondary antibody (Catalog No. 111-545-003; Jackson ImmunoResearch Laboratories, West Grove, PA, USA).
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