Somatostatin Antibody Staining Protocol for Immunohistochemistry

 

Description: Somatostatin is one of seven known polypeptide hormones produced in the pancreas. Somatostatin functions as an inhibitory hormone of the neuroendocrine system and is secreted by D-cells of the islets of Langerhans, the fundus and antrum of the stomach and in the upper small intestine. Somatostatin has been demonstrated in sympathetic nerves, mucosal cells, myenteric nerves of the gastrointestinal tract, salivary glands and in some parafollicular cells of the thyroid. This antibody labels D cells of mammalian pancreas. Cross-reactivity has been observed with somatostatin from a large number of mammalian species.

 

Primary Antibody

Name: Somatostatin Antibody

Clone: Rabbit anti-Human

Supplier: Dako

Catalog Number: A0566

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

Images: Search image

Additional Information:

Species Reactivity: Human, mouse, rat

Fixation: Formalin fixed paraffin sections

Positive Control: Pancreas (islet cells)

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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