Beta-Tubulin III Antibody Staining Protocol for Immunohistochemistry


Description: Beta-Tubulin III (Clone Tuj1). This antibody was raised against microtubules derived from rat brain and recognizes the peptide epitope, CEAQGPK. It is well characterized and highly reactive to neuron specific Class III β-tubulin (βIII). Tuj1 does not identify β-tubulin found in glial cells.


Primary Antibody

Name: Beta Tubulin III Antibody

Clone: Tuj1, Mouse monoclonal

Supplier: Covance

Catalog Number: MMS-435P

Dilution: 1:1200 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: Neuroal body and axons
Images: Search image

Additional Information:
Species Reactivity: Mouse, rat
Fixation: Formalin fixed paraffin sections
Positive Control: Brain (cerebral cortex)
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