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beta Amyloid, 1-16 (6E10) Antibody Staining Protocol for Immunohistochemistry
Description: This antibody is reactive to amino acid residues 1-16 of beta amyloid. The epitope lies within amino acids 3-8 of beta amyloid (EFRHDS). Beta amyloid forms are deposited in the CNS of patients with Alzheimer’s disease and Down’s syndrome. Signet’s beta amyloid antibody 6E10 reacts to the abnormally processed isoforms, as well as precursor forms.
Primary Antibody
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Name: Mouse Anti-beta Amyloid Antibody |
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Clone: 6E10 |
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Supplier: Covance |
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Catalog Number: SIG-39300 |
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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 Chromogen Substrate |
| Incubation Time/Temperature: 1-3 minutes/room temperature |
Counterstain
| Reagent: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Amyloid deposites |
| Images: Search image |
Additional Information:
| Species Reactivity: Human, mouse |
| Fixation: Formalin fixed paraffin sections |
| Positive Control: Alzheimer's brain |
| 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: