The Lightning-Link kit allows antibody conjugates to be set up in seconds by adding a solution of the protein to be labelled to a lyophilised mixture containing alkaline phosphatase (AP).
By circumventing the desalting or dialysis steps that commonly interrupt traditional protein conjugation procedures, Lightning-Link technology can be used to label small quantities of protein with 100 per cent recovery and without excessive dilution of the conjugate.
Upon dissolution of Lightning-Link mixture with a solution of the antibody (or other biomolecule to be labelled) chemicals in the mixture become activated.
This results in the directional coupling of the antibody to the enzyme label in a gentle and controlled process at near-neutral pH.
The hands-on time for the entire procedure is typically 20-30 seconds.
Lightning-Link allows the labelling of primary antibodies and other proteins and eliminates the need for secondary reagents in immunoassay procedures such as Western blotting, Elisa and immunocytochemistry.
Alkaline phosphatase (AP) is a hydrolyase enzyme that is frequently conjugated to antibodies for use in immunoassays.
AP has a lower catalytic rate than HRP, but has greater stability and maintains linear reaction kinetics for much longer.
PNPP is a popular colorimetric substrate for AP; the product of the reaction, p-nitrophenol, can be read at 405nM.
Fluorogenic substrates (e.g 4-methylumbelliferyl phosphate; MUP) may also be employed.
As well as AP, the company offers more than 40 available labels including enzymes, fluorescent proteins and dyes that span a variety of applications (WB, IH, FACS, Elisa, and Fret), colours and emission ranges.
