Thermo Scientific Lumi-Phos WB Substrate is a chemiluminescent substrate for sensitive detection of alkaline phosphatase activity from AP-conjugated antibodies and probes in Western blot detection.
Lumi-Phos WB Chemiluminescent Substrate overcomes the limitations posed by conventional chemiluminescent substrates for alkaline phosphatase (AP). Lumi-Phos WB Substrate provides Western blot detection of low-picogram (attomole) levels of protein targets, which is comparable in sensitivity to many ECL substrates for horseradish peroxidase (HRP). Lumi-Phos WB Substrate also produces less background noise than other commercially available alk-phos chemiluminescent substrates, providing a better signal:noise ratio and a clearer image when exposed to autoradiography film or CCD imagers. Because signal generation is immediate, there's no need to wait 15 to 30 minutes for a measurable signal. All of these benefits are combined with an affordable price.
- Alk-phos substrate – a chemiluminescent substrate for detection of alkaline phosphatase (AP or alk-phos) activity from antibodies and other Western blot probes
- Highly sensitive – detect low-picogram amounts of protein targets; detected 1.2pg (71 attomoles) of mouse IL-2 in example experiment
- Low background – provides high signal:noise ratios to yield clear blots that are easily documented by exposure to x-ray film or imagers
- Inexpensive – less costly to use than CDP-Star Substrate (based on 2008 U.S. list prices), and there is no need to purchase additional enhancers for nitrocellulose membranes
- Long signal duration – when conditions are optimized, blots generate usable signal for up to 8 hours (Em-max equals 440nm)
- Immediate strong signal – no more waiting 15 to 30 minutes for the signal to become strong enough to detect with imagers
- Ready to use – no mixing required with this one-component system
|Thermo Scientific Lumi-Phos Substrate provides high sensitivity and low background. Serial dilutions of recombinant mouse IL-2 were separated by 4-20% SDS-PAGE, then transferred to nitrocellulose membrane. The membranes were subsequently incubated in with rat anti-mouse IL-2 (1µg/mL), followed by AP-labeled goat anti-rat IgG (200ng/mL). The membranes were washed and then incubated in Lumi-Phos WB Substrate for five minutes before film exposure.
- Bruinsma, P., et al. (2004). J. Biol. Chem. 279, 39814-39823.
- Dhingra, A., et al. (2004). Proc. Nat. Acad. Sci. USA 101, 6315-6320.
- Tikhonov, I., et al. (2003). J. Virol. 77, 3157-3166.
Western blotting and detection technical guide
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