The Human beta Catenin (Total)ELISA Kit is a solid-phase sandwich Enzyme-Linked Immunosorbent Assay (ELISA) designed to detect and quantify the level of beta Catenin (Total) in fresh or frozen human cell lysates. Cross-reactivity has bee observed in mouse and rat cells. The assay recognizes both natural and recombinant beta Catenin (Total), independent of its phosphorylation state.
Principle of the method
A monoclonal capture antibody specific for beta Catenin (Total) has been coated onto the wells of the 96-well plate. During the first incubation, standards of known content and unknown samples are pipetted into the wells and the antigen binds to the immobilized (capture) antibody. After washing, a rabbit antibody specific for the target protein is added to the wells and serves as a detection antibody by binding to the immobilized protein captured during the first incubation. After washing, a horseradish peroxidase labeled anti-rabbit IgG is added. This binds to the detection antibody to complete the four member sandwich. After a third incubation and washing to remove all the unbound enzyme, a substrate solution (TMB) is added, which is acted upon by the bound enzyme to produce color. The intensity of this colored product is directly proportional to the concentration of target protein present in the original specimen and the optical density can be read on a standard microplate reader.
Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.
Beta-catenin, an adherens junction (AJ) protein, was originally identified as a component of cell-cell adhesion structures. AJs are necessary for the creation and maintenance of epithelial cell layers by regulating cell growth and adhesion between cells. Beta-catenin interacts with the cytoplasmic domain of E-cadherin and links E-cadherin to alpha-catenin, which in turn mediates anchorage of the E-cadherin complex to the cortical actin cytoskeleton. Studies show that Beta-catenin also binds to another cytoskeletal complex containing the adenomatous polyposis coli protein and microtubules, and interacts with several signaling pathways that include tyrosine kinases, phosphatases and Wnt/Wingless. The interplay between beta-catenin, cytoskeletal complexes and signaling pathways may regulate morphogenesis. Beta-catenin is expressed in several hair follicle cell types, basal and peripheral matrix cells, and cells of the outer and inner root sheats. A pathological role of beta-catenin has been identified in pilomatrixoma (PTR), medulloblastoma (MDB), colorectal cancer (CRC), ovarian cancer, and tumor development. In the nucleus, beta-catenin serves to co activate a family of Lef/Tcf transcription factors that stimulate transcription of target genes including those encoding cyclin D and c-myc that promote cell proliferation. The influence on cell proliferation is the molecular basis for the role of beta-catenin in tumorgenesis, specifically, solid tumors of the breast, colon, liver, lung, gastric, prostate, and skin.
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