Tau Antibody Staining Protocol for Immunohistochemistry


Description: The brain tissues from patients with Alzheimer's disease are characterized by an abundance of neurofibrillary tangles, neuropil threads and abnormal neurites in senile plaques. Tangles represent dense accumulations of ultrastructurally distinct paired helical filaments whose major component is a microtubule-associated tau protein. This antibody raised against the bovine tau protein, crossreacts with the phosphorylated form of human tau protein enabling the conformational dependent epitope found only in the neurofibrillary components in Alzheimer's disease cases in humans.


Primary Antibody

Name: Tau Antibody

Clone: Tau-2, Mouse anti-Human

Supplier: Novocastra

Catalog Number: NCL-Tau-2

Dilution: 1:100 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: Cytoplasmic (neurofibrillary tangles, neuropil threats and neuritis surrounding senile plaques)
Images: Search image

Additional Information:
Species Reactivity: Human
Fixation: Formalin fixed paraffin sections
Positive Control: Brain tissue with Alzheimer’s disease
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