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Beta-Amyloid Antibody Staining Protocol for Immunohistochemistry
Description: Beta amyloid is an extracellular filamentous protein deposit found in the brain. It is the major protein component of amyloid cores and neuritic plaques and is also found as a deposit in neurofibrillary tangels. In man, Alzheimer's disease is the most common cause of senile dementia and is characterized by abnormal filamentous protein deposits in the brain. Beta amyloid deposits are also detected in Lewy body dementia, Down's syndrome, amyloidosis (Dutch type) and in the Guam Parkinson-Dementia complex.
Primary Antibody
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Name: Beta Amyloid Antibody |
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Clone: 6F/3D, Mouse Anti-Human |
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Supplier: Novocastra Labs |
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Catalog Number: NCL-B-Amyloid |
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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: 60min/room temperature |
Antigen Retrieval
| Device: Staining jar or other device |
| Buffer/pH value: Formic acid/pH 1.6-2.0 |
| Heat/Cool Temperature: Room temperature |
| Heat/Cool Time: Incubate for 10 - 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: Positive staining can be observed in senile plaque cores, plaque periphery and diffuse plaques. In some cases of Alzheimer’s disease staining can be observed in vessel walls and in extracellular neurofibrillary tangles. |
| Images: Search image |
Additional Information:
| Tissue Type: Brain tissue with Alzheimer’s disease, Lewy body dementia, Down's syndrome, amyloidosis (Dutch type) and in the Guam Parkinson-Dementia complex. |
| Fixation: Formalin-fixed paraffin sections |
| Positive Control: Brain tissue with Alzheimer’s disease, Lewy body dementia, Down's syndrome, amyloidosis (Dutch type) and in the Guam Parkinson-Dementia complex. |
| 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: