Novex AP Chemiluminescent Substrate should be combined with Nitro Block II for use with nitrocellulose membranes. The emitted light is stable up to 24-96 hours allowing for multiple exposures.Ĭhoose from a standalone substrate, Novex AP Chemiluminescent Substrate or the WesternBreeze Chemiluminescent Immunodetection kits which contains all solutions necessary for your application including blocking solutions, primary antibody diluent, ready-to-use secondary antibody solution, ready-to-use chemiluminescent substrate, wash solutions, incubation trays, pre-cut filter papers, polyester sheet for even substrate development on the membrane. With simply writing on the pre-stained marker, the position of the. This eBook explores these solutions in detail and the advantages they offer researchers conducting western blotting in the lab. LuminolPen, HRP System is a perfect tool for chemiluminescent detection in Western blot. Explore enzyme-based detection systems for ELISA, immunohistochemistry, and Western blotting including OPD, TMB, BCIP, pNPP, fast red, ABTS, DAB, chemiluminescence, and many more. Light emission occurs only during the enzyme-substrate reaction therefore, once the substrate in proximity to the enzyme is exhausted, signal output ceases. The Amersham ImageQuant 800 GxP biomolecular imagers, developed by Cytiva, offer many benefits for chemiluminescence and fluorescence detection, including enhanced sensitivity, resolution, and data security. This includes substrates for staining and labeling of actin microfilaments and the detection of cholesterol and lipoproteins in cells. The choice of substrate for chemiluminescent Western blotting is determined by which reported enzyme is chosen. The two commonly used enzyme reporters are HRP and AP. Overview of the western blotting procedure. CDP-Star is dephosphorylated by AP to yield meta-stable dioxetane phenolate anion intermediate that decomposes and emits light with a maximum intensity at a wavelength of 475 nm. Chemiluminescent detection systems take advantage of energy released in reporter enzyme-substrate reactions in the form of light to be the recorded signal. The protein of interest is probed with specific antibodies and detected through a number of means, commonly chemiluminescence. For western blot detection based on alkaline phosphatase (AP), 86 kDa, we offer our CDP-Star substrate that is designed to deliver picogram level sensitivity and is compatible with both traditional x-ray film and CCD-based imaging. The chemiluminescence method is mainly used for detection due to its high sensitivity and ease of manipulation, but it is unsuitable for detailed analyses because it cannot be used to detect multiple proteins simultaneously.
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