ELISA: 0.05 μg/mL. Western Blot: 0.5 μg/mL for HRPO/ECL detection. Recommended blocking buffer: Casein/Tween 20 based blocking and blot incubationbuffer. Immunoprecipitation: 1-10 μg per 10^6 vanadate treated A431 cells. Immunocytochemistry: 0.1-1 μg/mL. Immunohistochemistry on Paraffin Sections. Included Positive Control: Cell lysate from untreated SW480 cells (for details seeProtocols). Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
Protocol
Positive Control: Cell lysate from untreated SW480 cells. Formulation: Lyophilized Cell Lysate from Serum starved SW480 cells. Stability: Reconstitute by addition of 200 µl H2O. After complete solubilization add 200 µl 2xSDS-PAGE sample buffer, mix and incubate at 90°C for 5 min. Application: The Positive Control Cell Lysate is recommended for immunoblot applications. Use 20µlmolecular weight marker per lane. Note: Use BSA based blot incubation buffers. Milk,Casein and Blotto might interfere with antibody - antigen interaction.
Restrictions
For Research Use only
Reconstitution
Restore with 1 mL H2O (15 min, RT).
Buffer
1 mL2 x PBS containing 0.09 % Sodium Azide, PEG and Sucrose
Preservative
Sodium azide
Precaution of Use
WARNING: Reagents contain sodium azide. Sodium azide is very toxic if ingested or inhaled. Avoid contact with skin, eyes, or clothing. Wear eye or face protection when handling. If skin or eye contact occurs, wash with copious amounts of water. If ingested or inhaled, contact a physician immediately. Sodium azide yields toxic hydrazoic acid under acidic conditions. Dilute azide-containing compounds in running water before discarding to avoid accumulation of potentially explosive deposits in lead or copper plumbing.
Storage
-20 °C/-80 °C
Storage Comment
Store lyophilized (preferably in a desiccator) at -20 °C and reconstituted (aliquote andfreeze in liquid nitrogen) at -80 °C. Avoid repeated freezing and thawing. Thaw aliquots at 37 °C. Thawed aliquots may be stored at 4 °C up to 3 months. Shelf life: one year from despatch.
The α-, β- and γ-catenins are cytoplasmic proteins mediating the interaction of Ca2+-dependent transmembrane adhesion molecules (cadherins) with the cytoskeletal network. The direct interaction of β-catenin with the cytoplasmic domain of cadherins plays a crucial role for cell-cell adhesion and signal transmission between neighbouring cells. Recent studies indicate that β-catenin may also play a role in tumorigenesis since it forms complexes with the tumor suppressor gene product APC. β-catenin directly interacts and constitutively activates transcription factors of the TCF/LEF gene family. Thus it is proposed that β-catenin plays a dual role not only in the maintainance and regulation of cell-cell interactions but also in the regulation of gene activity.Synonyms: Beta-catenin, CTNNB, CTNNB1