Beta Catenin Antibody Staining Protocol for Immunohistochemistry


Description: The catenins (α, β, γ, δ) are ubiquitously expressed, cytoplasmic proteins that associate with E-cadherin at cellular junctions. α-catenin also associates with P-cadherin and N-cadherin. β-catenin co-immunoprecipitates with APC. γ-catenin associates with both N-cadherin and E-cadherin and is a major component of desmosomes, where it is found complexed to desmoglein. δ-catenin interacts with Presenilin 1. A hemizygous deletion of the gene encoding δ-catenin correlates with mental retardation in cri-du-chat syndrome. Armadillo Repeat gene deleted in Velo-Cardio-Facial syndrome (ARVCF) is also a member of the catenin family that may function as a nuclear protein.


Primary Antibody

Name: Beta Catenin Antibody

Clone: Goat anti-beta Catenin

Supplier: Santa Cruz Biotechnology

Catalog Number: sc-1496

Dilution: 1:200 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: 60min/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: Membrane/Cytoplasmic (can translocated to nucleus under certain conditions)
Images: Search image

Additional Information:
Tissue Type: Intestine
Fixation: Formalin fixed paraffin sections
Positive Control: Intestine
Negative Control: Omit primary antibody, isotype control or 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