Immunohistochemistry was performed as described previously30 (link). Briefly, 3–7-day old brains were dissected in ice-cold PBS, fixed in 4% paraformaldehyde, and blocked in PBST (PBS and 0.3% Triton X-100) with 5% normal donkey serum (Jackson Laboratory) before staining. Brains were then incubated with 1:1,000 rabbit anti-GFP (Invitrogen) and 1:50 mouse anti-nc82 (Developmental Studies Hybridoma Bank) antibodies overnight at 4 °C and washed five times in PBST. Brains were then incubated with 1:1,000 Alexa 568 anti-rabbit (Life Technologies) and 1:1,000 Alexa 633 anti-mouse (Life Technologies) antibodies for 4 h at room temperature before washing five times in PBST and coverslip mounting in Vectashield (Vector Labs). Images were taken at × 40 magnification on a Leica SP5 confocal microscope at 0.5 mm intervals and reassembled for display as maximum projections using Fiji56 (link).
Alexa 633 anti mouse
Manufactured by Thermo Fisher Scientific
Alexa Fluor 633 anti-mouse is a fluorescently labeled secondary antibody used for detection and visualization in various immunological techniques. It binds to mouse primary antibodies, allowing for sensitive and specific detection of target proteins or cells in samples.
Automatically generated - may contain errors
Lab products found in correlation
Normal donkey serum (nds), by Jackson ImmunoResearch (1 mentions)
Vectashield, by Vector Laboratories (1 mentions)
Sp5 confocal microscope, by Leica (1 mentions)
Rabbit anti gfp, by Thermo Fisher Scientific (1 mentions)
Mouse anti nc82, by Developmental Studies Hybridoma Bank (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Monoclonal mouse anti occludin, by Zymo Research (1 mentions)
2 protocols using alexa 633 anti mouse
1
Immunohistochemistry of Drosophila Brains
2
3D Co-culture and Tight Junction Staining
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Human macrophages or dendritic cells were pre-stained with 2 mg/ml of 6-carboxyfluorescein diacetate before they were used in the co-culture. The 3D co-culture was fixed with 3% paraformaldehyde. Tight junctions were stained with monoclonal mouse anti-occludin (Zymed, San Francisco, CA; 1:200) and Alexa633 anti-mouse (Life Technologies, Darmstadt, Germany; 1:500), nuclei were stained with DAPI (Life Technologies, Darmstadt, Germany). Detailed protocol is described in SI.
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