Tyrosine Hydroxylase (TH) Antibody Staining Protocol for Immunohistochemistry


Description: Tyrosine hydroxylase is the first enzyme in catecholamine (CA) biosynthesis and catalyses the conversion of L-tyrosine to L-DOPA. Tyrosine hydroxylase is , therefore, a useful marker of all CA neurons and allows their localization in different areas of the brain.


Primary Antibody

Name: Tyrosine Hydroxylase (TH) Antibody

Clone: Rabbit polyclonal

Supplier: Millipore

Catalog Number: AB152

Dilution: 1:100 - 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 Chromogen Substrate
Incubation Time/Temperature: 1-3 minutes/room temperature

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Staining Pattern: Cytoplasmic of dopamine neurons
Images: Search image

Additional Information:
Species Reactivity: Human, mouse, rat, ferret, feline, Aplysia (marine mollusc).
Fixation: Formalin fixed paraffin sections or formalin fixed frozen sections
Positive Control: Brain (striatum, substantia nigra) dopamine neurons
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