Synaptophysin Antibody Staining Protocol for Immunohistochemistry

 

Description: It recognizes a protein of 38kDa, identified as synaptophysin and labels normal neuroendocrine cells of human adrenal medulla, carotid body, skin, pituitary gland, thyroid, lung, pancreas, gastrointestinal mucosa, Paneth’s cells in the gastrointestinal tract and of gastric parietal cells. Neurons in the brain, spinal cord, and retina are also labeled. Anti-synaptophysin antibody reacts with neuroendocrine neoplasms of neural as well as epithelial types e.g. neuroblastomas, ganglioneuroblastomas, ganglioneuromas, pheo-chromocytomas, chromaffin, and non-chromaffin paragangliomas.

 

Primary Antibody

Name: Synaptophysin Antibody

Clone: Rabbit anti-Human

Supplier: Lab Vision

Catalog Number: RB-9044-P

Dilution: 1:500 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

Counterstain

Reagent: Mayer's Hematoxylin

Staining Time: 30 seconds

Results:

Staining Pattern: Cytoplasmic/membrane

Images: Search image

Additional Information:

Species Reactivity: Human, rat

Fixation: Formalin fixed paraffin sections

Positive Control: Pancreas or pheochromocytoma

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

 

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