beta catenin antibody,anti-β-catenin单克隆抗体

PreparationThis antibody was produced from a hybridoma resulting from the fusion of a mousemyeloma with B cells obtained from a mouse immunized with purified, E. coli-derivedrecombinant human beta-Catenin (GenBank accession # NM_001904). The IgG fraction ofthe tissue culture supernatant was purified by Protein G chromatography. Beta-Catenin is acadherin-associated protein that is involved in the regulation of cell adhesion. It is alsoinvolved in the canonical Wnt signal transduction pathway.FormulationLyophilized from a 0.2 μm filtered solution in phosphate-buffered saline (PBS) with5% trehalose.ReconstitutionReconstitute with sterile PBS to achieve a final concentration of 500 μg/mL.StorageLyophilized samples are stable for twelve months from date of receipt when stored at -20° Cto -70° C. Upon reconstitution, the antibody can be stored at 2° - 8° C for 1 month withoutdetectable loss of activity. Reconstituted antibody can also be aliquotted and stored frozenat -20° C to -70° C in a manual defrost freezer for six months without detectable loss ofactivity. Avoid repeated freeze-thaw cycles.SpecificityThe antibody was selected for its ability to detect endogenous human, mouse and ratbeta-Catenin in Western blots.ApplicationWestern blot - An antibody concentration of 2.0 μg/mL is recommended.Optimal dilutions should be determined by the individual laboratory.Protocols for Immunoblotting:Western blottingBlotting buffer Blocking Solution Antibody Solution25 mM Tris, pH 7.40.15 M NaCl0.1% Tween 202% nonfat dry milk inblotting bufferAdjust pH to 7.42% nonfat dry milk inblotting bufferAdjust pH to 7.41. Transfer the electrophoresed proteins to Immobilon-P membrane (Millipore)and incubate the membrane for 1 hour at room temperature in BlockingSolution.2. Incubate the membrane overnight at 4° C in antibody solution containing 2.0 μg/mLmouse anti-human/mouse/rat beta Catenin.3. Wash the membrane at room temperature for 30 minutes with 3 or morechanges of Blotting Buffer. Changing the membrane containers often reducesbackground.4. Incubate the membrane at room temperature for 1 hour in Antibody Solutioncontaining a 1:2,000 dilution of HRP-conjugated sheep anti-mouse IgG(Amersham).5. Wash the membrane for 1 hour with 5 or more changes of Blotting Buffer.6. Detect with WesternGlo Chemuluminescent Detection Substrate (R&DCatalog # AR004).Cell lysates for Western blottings: To prepare total cell lysates, cells aresolubilized in hot 2x SDS gel sample buffer (20 mM dithiothreitol, 6% SDS, 0.25 MTris, pH 6.8, 10% glycerol, 10 mM NaF and bromophenyl blue) at2 x 106 - 1 x 107 cells per mL. The extracts are heated in a boiling water bath for5 minutes and then sonicated with a probe sonicator with 3 - 4 bursts of5 - 10 seconds each. Samples are diluted with 1x SDS sample buffer to thedesired concentration.