Beta-galactosidase Alpha Complementation Kit - (1 kit)
Beta-galactosidase Alpha Complementation Kit - (1 kit)
Beta-galactosidase Alpha Complementation Kit - (1 kit)

Beta-galactosidase Alpha Complementation Kit - (1 kit)

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A Beta-galactosidase Alpha complementation kit consisting of enzyme donor, enzyme acceptor, their respective stabilization buffers, a Beta-galactosidase chromogenic substrate and detailed instructions. Kit can be used to develop alpha complementation-based assays (cloned enzyme donor immunoassays).

 

Product Data Sheet Download PDF
Catalog # K2010002
Content
  • Enzyme Acceptor: 2.5 mg lyophilized powder
  • Enzyme Donor: 0.25 mg lyophilized powder
  • Enzyme Donor Stabilization Buffer: 25 mL
  • Enzyme Acceptor Stabilization Buffer: 25 mL
  • Beta-galactosidase Chromogenic Substrate: 25 mg
  • Beta-galactosidase Chromogenic Substrate Buffer: 25 mL
  • Detailed Instructions
Storage -20°C. Avoid repeated freeze/thaw cycles.
Supplied as Research Kit
Buffer Contains stabilization buffers
Purity All enzymes are > 95% pure
Application Alpha complementation kit, cloned enzyme donor immunoassays, ELISA.
Keywords Beta-galactosidase Alpha Peptide, Beta-galactosidase Enzyme Donor, Beta-galactosidase Omega Domain, Enzyme Acceptor, Alpha Complementation, LacZ.
Enzymes Source Recombinant
Related products
  • Beta-galactosidase Enzyme Donor (Alpha Peptide)
  • Stabilization Buffer for Enzyme Acceptor
  • Beta-galactosidase Enzyme Acceptor (Omega Domain)
  • Beta-galactosidase Enzyme Acceptor Stabilization Buffer
  • Beta-galactosidase Chromogenic Substrate

About Beta-galactosidase Alpha Complementation

Beta-galactosidase Alpha-Complementation is a biochemical phenomenon first documented by Agnes Ullmann, while working in the lab of François Jacob and Jacques Monod. By means of molecular cloning, the native E. coli β-galactosidase enzyme can be split in two inactive fragments of different sizes. The smaller fragment, known as the alpha-peptide or enzyme donor, is about 100 amino residues in length and is inactive on its own (incapable of hydrolyzing a β-galactosidase substrate). The larger fragment, known as the omega fragment or enzyme acceptor, is about 900 amino residues in length and is also inactive on its own. Upon mixing the enzyme donor with the enzyme acceptor, the β-galactosidase enzyme is reconstituted and is now capable of hydrolyzing colorimetric substrates such as ONPG.

Both enzyme donor and enzyme acceptor can be cloned and expressed in special E. coli strains to yield highly pure, zero-background enzyme fragments (i.e. an enzyme donor and enzyme acceptor without measurable catalytic activities, when assayed individually). Interestingly, it was discovered that various analytes can be conjugated to the enzyme donor moiety and the enzyme donor-enzyme acceptor association modulated by an analyte-binding molecule (such as an antibody). As a result, an alpha-complementation-based assay can be developed.

References

  1. Kras, E. (2019). Beta-galactosidase: properties, structure and functions. New York: Nova Science Publishers.

  2. Arndt, T. (2017). Cloned Enzyme Donor Immunoassay. Lexikon Der Medizinischen Laboratoriumsdiagnostik, 1–2.

  3. Jeon, S. I., Yang, X., and Andrade, J. D. (2004). Modeling of homogeneous cloned enzyme donor immunoassay. Analytical Biochemistry, 333(1), 136–147.

  4. Tachi, T., Kaji, N., Tokeshi, M., and Baba, Y. (2009). Microchip-based Homogeneous Immunoassay Using a Cloned Enzyme Donor. Analytical Sciences, 25(2), 149–151.

  5. Khanna, P. L., and Worthy, T. E. (1993). CEDIA: A Recombinant-Based Homogeneous Enzyme Immunoassay.