A key downstream effector in the Wnt signaling pathway is b-catenin (1). It is implicated in two major biological processes of vertebrates: early embryonic development (2) and tumorigenesis (3). Casein kinase 1 phosphorylates b-catenin on Ser45. This phosphorylation event primes b-catenin for subsequent phosphorylation by GSK-3 (4-6). GSK-3b destabilizes b-catenin by phosphorylating it at Ser33, 37 and Thr41 (7). Mutations in these phosphorylation sites which result in the stabilization of b-catenin protein levels have been found in many tumor cell lines (8).
This ELISA Kit is a solid phase sandwich enzyme-linked im munosorbent assay (ELISA) that detects endogenous levels of total beta-catenin protein (phosphorylated and non-phosphorylated). A beta-Catenin antibody has been coated onto the microwells. After incubation with cell lysates, both phospho- and nonphospho-beta-catenin proteins are captured by the coated antibody antibody. Following extensive washing, beta-Catenin mouse monoclonal antibody is added to detect both the captured phospho- and nonphospho-beta-catenin protein. An anti-mouse IgG, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color color. The magnitude of optical density for this developed color is proportional to the quantity of total beta-catenin protein.
1. Microtiter Plate, Catenin antibody-coated microtiter, 1x96 wells.
2. Beta-Catenin Detection Ab, 1x11ml
3. IgG (HRP) Ab, 1x11ml
4. TMB Substrate, 1x 11ml
5. STOP Solution, 1x11ml
6. Wash Buffer (20X), 1x25ml
7. Sample Diluent, 1x25ml.
8. Cell Lysis Buffer, 1x15ml
Storage and Stability:
Store all kit components at 4 degrees C except Cell Lysis Buffer. Store Cell Lysis Buffer at -20 degrees C.
Stable for 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.