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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
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Name: Tau Antibody |
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Clone: Tau-2, Mouse anti-Human |
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Supplier: Novocastra |
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Catalog Number: NCL-Tau-2 |
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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. |
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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 (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 |
References: