3rd instar larval brains were dissected out in 1 × PBS, fixed in 4% paraformaldehyde. Staining was performed following the standard protocol by Daul et al. (2010) . Brains were finally mounted using Vectashield Dapi (H-1200) (Vector Laboratories) mounting media. The primary antibody was Mira (1:200). Pictures were taken using Olympus FV-1000 confocal microscope.
Vectashield dapi h 1200
Manufactured by Vector Laboratories
Vectashield Dapi (H-1200) is a mounting medium designed for fluorescence microscopy. It contains the nuclear counterstain 4',6-diamidino-2-phenylindole (DAPI), which binds to DNA and emits blue fluorescence when excited by ultraviolet light. This product is used to visualize and label cell nuclei in fixed and permeabilized samples.
Automatically generated - may contain errors
Lab products found in correlation
Fv1000 confocal microscope, by Olympus (4 mentions)
Sc 12918 r, by Santa Cruz Biotechnology (2 mentions)
Duo92002, by Merck Group (1 mentions)
Duo92004, by Merck Group (1 mentions)
Duo92008 3, by Merck Group (1 mentions)
Confocal microscope, by Olympus (1 mentions)
Fluoview fv1000, by Olympus (1 mentions)
Teflon printed 12 well slides, by Electron Microscopy Sciences (1 mentions)
Lsm 710 meta laser confocal microscope, by Zeiss (1 mentions)
Elyra super resolution microscope, by Zeiss (1 mentions)
7 protocols using vectashield dapi h 1200
1
3rd Instar Larval Brain Staining
2
Drosophila Brain Immunohistochemistry
3
Immunostaining and Duolink Assay for Protein Interactions
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Cells on coverslips were fixed using 4% paraformaldehyde (Electron Microscopy Sciences #15710). Cell membranes were permeabilized using 0.2% Triton X-100 (Thermo Fisher Scientific #9002–93-1). Cells were immuno-stained with anti-V5 and anti-Flag antibodies followed by duolink assay (DUO92008-3, DUO92004, DUO92002, Sigma-Aldrich) according to the manufacturer’s instructions. Coverslips were mounted with VectaShield/DAPI (H-1200, Vector Laboratories) on microscopy slides and analyzed using Olympus confocal microscope and Olympus Fluoview FV1000 software (https://www.olympus-lifescience.com/en/downloads/detail-iframe/?0[downloads][id]=847249651 ).
4
Enzyme-Linked Immunosorbent Assay for Perkinsus marinus
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Teflon printed 12-well slides (Electron Microscopy Sciences, Hatfield, PA) were coated with 3×104P. marinus parasites in 1× PBS/1%BSA/well, air-dried and stored at −80°C until use. Slides were thawed for 30 min at room temperature and then blocked with 1xPBS/1% BSA for 30 min at 37°C. Serum samples at various dilutions (two-fold dilution series starting at 1∶20) were added to the parasite-coated wells and incubated for 1 h at 37°C. Slides were washed three times with 1xPBS, incubated with FITC-labeled F (ab’)2 goat anti-mouse IgM, IgG, or IgA (Southern Biotechnologies, Birmingham, AL) at a 1∶40 dilution in 1xPBS/0.1% Evans blue for 30 minutes at 37°C. Slides were washed, air dried, and mounted with VECTASHIELD-DAPI (H-1200, Vector laboratories, Burlingame, CA).
5
Immunofluorescence Assay for TDP-43 Localization
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For these assays, 293 T cells were transfected with or without plasmids and then fixed with 3.7% paraformaldehyde in PBS at RT for 15 min at 24 h (for GFP-TDP-43-IIPLD experiments) or 48 h post-transfection. The fixed cells were incubated with a TDP-43 or lamin A/C or His antibody at 4 °C overnight, followed by incubation with a secondary antibody conjugated with a fluorescent dye (Molecular Probes). The staining cells were gently fixed with 3.7% paraformaldehyde for 2 min before mounting. Slides were mounted using Vectashield DAPI H-1200 (Vector Laboratories). Cellular fluorescence images were obtained from a single optical section using an LSM710 META laser confocal microscope (Zeiss) or ELYRA super-resolution microscope (Zeiss)33 (link).
6
3rd Instar Larval Brain Staining
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3rd instar larval brains were dissected out in 1 × PBS, fixed in 4% paraformaldehyde. Staining was performed following the standard protocol by Daul et al. (2010) . Brains were finally mounted using Vectashield Dapi (H-1200) (Vector Laboratories) mounting media. The primary antibody was Mira (1:200). Pictures were taken using Olympus FV-1000 confocal microscope.
7
Drosophila Brain Immunohistochemistry
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Adult Drosophila brains were dissected out in 1 × PBS by carefully removing the cuticle surrounding the brain. The brain was fixed with 4% paraformaldehyde in 1 × PBS for 90 min by end over end mixing. The fixed brains were washed with 0.5% Triton X-100 in 1 × PBS for 30 min to 1 hr. Brains were then incubated for 2 days at 4°C in primary antibody in 0.5% Triton X-100 in 1 × PBS with 10% BSA (antibody dilution buffer). After primary antibody incubation, brains were washed with 0.5% Triton X-100 in 1 × PBS 3 times, 15 min each. The secondary antibody was also diluted in antibody dilution buffer and incubated overnight at 4°C. Finally, the brains were washed 3 times and added to Vectashield Dapi (H-1200) (Vector Laboratories) mounting media and kept for 2 days in dark at 4°C. The brains were finally mounted using Vectashield Dapi (H-1200) mounting media. Primary antibodies are: Mira (Miranda) (1:200) (gift from Dr. Chengyu Lee) and phospho-Cdk5 (1:200) (sc-12918-R) (Santa Cruz). Pictures were taken using Olympus FV-1000 confocal microscope.
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