Synaptophysin Antibody Staining Protocol for Immunohistochemistry


Description: Synaptophysin is an integral membrane glycoprotein present in presynaptic vesicles in almost all neurons. This antibody reacts with a hydrophilic peptide sequence selected from the deduced amino acid sequence of synaptophysin. This antibody labels neuroendoctine cells of the human adrenal medulla, carotid body, skin, pituitary gland, thyroid, lung, pancreas, and gastrointestinal mucosa. Also labelled are neurons of the brain, spinal cord and retina. Neuroendrocrine neoplasms including neuroblastomas, ganglioneuroblastomas, ganglioneuromas, pheochromocytomas, chromaffin, and non-chromaffin paragangliomas are labelled. Epithelial neuroendocrine neoplasms of the pituitary, pancreas, thyroid, lungs, gastointestinal tract and skin are also reactive.


Primary Antibody

Name: Synaptophysin Antibody

Clone: Rabbit anti-Human

Supplier: Dako

Catalog Number: A0010

Dilution: 1:200 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed.

Incubation Time/Temp: 60 min/room temperature

Antigen Retrieval
Device: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102)
Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100)
Heat/Cool Temperature: 95-100 ºC/room temperature
Heat/Cool Time: 20 minutes/20 minutes

Detection Methods
Standard Method: ABC Method or LSAB Method
Enhanced Method: Polymeric Methods

Chromogen Substrate
Reagent: DAB
Incubation Time/Temperature: 1-3 minutes/room temperature

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Staining Pattern: Membrane
Images: Search image

Additional Information:
Species Reactivity: Human
Fixation: Formalin fixed paraffin sections
Positive Control: Brain
Negative Control: Omit primary antibody, isotype control, absorption control
Blocking: 2-5% normal serum to reduce unspecific background staining; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary