Immunofluorescent analysis of Catenin beta (green) in HeLa cells either left untreated (left panel) or treated with 10uM GSK-3 Inhibitor X for 20 hours (right panel). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with a Catenin beta polyclonal antibody (Product # PA5-16762) at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-rabbit IgG secondary antibody (Product # 35552) at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight 554 Phalloidin (Product # 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan or a ToxInsight Instrument at 20X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A synthetic peptide from the C-terminus of the beta-catenin protein|
|Storage buffer||PBS, pH 7.6, with 0.2% BSA|
|Contains||<0.1% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:250-1:500|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||2 µg/ml|
|Western Blot (WB)||See 1 publications below|
This antibody reacts with the C-terminal portion of beta-Catenin.
For staining of formalin-fixed, paraffin-embedded tissues, it is recommended to boil tissue sections in 10mM citrate buffer, pH 6.0 for 10-20 min followed by cooling at room temperature for 20 min. Recommended positive controls are MCF-7 cells and breast carcinoma tissue.
This antibody was orginally validated as part of a Thermo Scientific Cellomics High Content Screening Kit. The antibody sold separately may have slightly different performance and may need to be further optimized for the best results.
The catenins (alpha, beta and gamma) are ubiquitously expressed, cytoplasmic proteins associated with E-cadherin at cellular junctions. beta-catenin also binds to N-cadherin and co-immunoprecipitates with APC. Cadherin/catenin complexes are linked to the cytoskeleton via a direct association between alpha-actinin and alpha-catenin. Increases tyrosine phosphorylation can disrupt catenin-cadherin complexes, influencing cellular adhesion.
IP-MS enrichment of CTNNB1 (LFQ intensity): CTNNB1 was enriched 377-fold from HCT116 lysate compared to background proteins, using the optimized IP-MS workflow with Pierce MS-Compatible Magnetic IP Kit protein A/G (Product # 90409) and CTNNB1 antibody (Product # PA5-16762). STRING database was used to identify the protein interactor list. See more information on IP-MS verification of antibody selectivity. IP-MS validation info.
Inhibition of discoidin domain receptor 2-mediated lung cancer cells progression by gold nanoparticle-aptamer-assisted delivery of peptides containing transmembrane-juxtamembrane 1/2 domain.
PA5-16762 was used in western blot to use gold nanoparticle-DNA aptamer conjugated to the functional domain of DDR2 to target lung cancer cells
|Kim D,Yeom JH,Lee B,Lee K,Bae J,Rhee S||Biochemical and biophysical research communications (464:392)||2015|