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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,
Primary Antibody
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Name: Synaptophysin Antibody |
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Clone: Rabbit anti-Human |
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Supplier: Dako |
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Catalog Number: A0010 |
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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. |
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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 |
Counterstain
| Reagent: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| 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 |
References: