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Method Article
This protocol demonstrates how to perform immunohistochemistry on dissected Drosophila larva.
Before you start
Immunohistochemistry
Mounting
Note : Mount in glycerol if samples will be imaged immediately. Mount in Prolong Gold if samples will be stored before imaging (for more than a week) or if samples must be imaged multiple times. To use, place a bottle of prolong in 65ºC water bath for 2-3 minutes. Take an aliquot of Prolong Gold and keep in on a heat-block at 45-50º C.
Immunohistochemistry (IHC) is vital for the study of NMJ biology because it enables visualization of the NMJ. This is accomplished by using antibodies that recognize the neuronal membrane (e.g., HRP), the presynapse (e.g., CSP, SYT), and/or the postsynapse (e.g., DLG). Signaling molecules, structural proteins, or novel proteins of interest can also be stained. Then, genes can be mutated or missexpressed, and IHC can detect perturbation of synaptic structure and/or neuronal signaling.
The NMJ o...
Name | Company | Catalog Number | Comments |
Stereomicroscope “Stemi” 2000 | Carl Zeiss, Inc. | 495101-9804-000 | |
Light Source KL 1500 LCD | Carl Zeiss, Inc. | 000000-1063-181 | |
Dumont SS Forceps | Fine Science Tools | 11200-33 | |
Dumont #5 Forceps | Fine Science Tools | 11252-20 | |
Adams™ Nutator Mixer | BD Biosciences | 421105 | |
No.1 Precision knife | X-Acto | 3201 | |
No. 16 Blades | X-Acto | 216 | |
ProLong Gold Antifade reagent | Invitrogen | 36930 |
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