Amyloid Precursor Protein Antibody Staining Protocol for Immunohistochemistry

 

Description: Different isoforms of amyloid precursor protein (APP) exist as a result of alternative splicing of a 19-exon gene. APP695, APP751 and APP770 are the predominant APP isoforms. In brain neuronal tissue APP695 is predominantly expressed while APP751 and APP770 are found in other tissues. APP, a transmembrane glycoprotein, plays a role in intracellular signalling by regulating cell-cell or cell-substrate interactions. It is involved in synaptogenesis. Normal synthesis of APP by nerve cells involves cleavage and release from the cell surface, but abnormal cleavage of APP results in a fragment called beta amyloid peptide that aggregates and forms plaques. These plaques are associated with Alzheimer’s Disease.

 

Primary Antibody

Name: Amyloid Precursor Protein Antibody

Clone: 3G12, Mouse Anti-Human

Supplier: Novocastra Labs

Catalog Number: NCL-APP-228

Dilution: 1:25 - 1:50

Incubation Time/Temp: 60 min/room temperature

Antigen Retrieval

Device: Steamer

Buffer/pH value: EDTA/pH 8.0

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: Gill's Hematoxylin or Mayer's Hematoxylin

Staining Time: 30 seconds

Results:

Staining Pattern: Neurofibrillary tangles and senile plaques

Images: Search image

Additional Information:

Tissue Type: Human brain, Alzheimer’s disease

Fixation: Formalin-fixed paraffin sections

Positive Control: Human brain, 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

 

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