Lumi-LightPLUS Western Blotting Substrate
ROCHE/12015196001 - pkg of 100 mL (1,000 cm2 membrane)
Synonym: western blotting substrate
Product Type: Chemical
Detect as little as 5 pg. Different amounts of Β-galactosidase were separated by SDS-PAGE, 6% acrylamide, and then blotted onto a PVDF membrane. After incubation with anti-Β-galactosidase monoclonal antibody (3 μg/mL), horseradish peroxidase-labeled mouse antibody (50 mU/mL) was used as a secondary detection reagent. Detection was performed for 10 minutes using the Lumi-Imager Instrument.
Time-response-curve of a Lumi-Light signal (in Boehringer Light Units) from a typical western blot, analyzed on the Lumi-Imager Instrument.
| manufacturer/tradename | Roche |
| packaging | pkg of 100 mL (1,000 cm2 membrane) |
| Quality Level | 100  |
| shipped in | wet ice |
| storage temp. | 2-8°C |
| Application: | Lumi-LightPLUS Western Blotting Substrate has been used in western blotting. |
| Application: | Lumi-LightPLUS Western Blotting Substrate is for the chemiluminescent detection of antigens blotted on membranes using horseradish peroxidase-conjugated secondary antibodies. It can be used for very sensitive detection of any antigen blotted on a PVDF Western Blotting Membrane or nitrocellulose membrane. Lumi-LightPLUS Western Blotting Substrate is suited for high-sensitivity western blotting, especially when quantification is required. The chemiluminescent signal persists for more than 9 hours, allowing multiple exposures to be taken. |
| Features and Benefits: | • Detect rare proteins. Detect antigen in the range of 1 - 5 pg. • Take multiple exposures. The signal is stable for more than nine hours after substrate addition. • Save sample material. Lumi-LightPLUS substrate detects less than 5 pg-amounts of antigen. • Save valuable resources. Dilute primary antibody 10- to 100-fold more than with colorimetric detection systems. • Benefit from easy preparation. Simply mix the two Lumi-Light components in a 1:1 ratio. |
| General description: | As the technology for quantifying chemiluminescence advances, the substrates for chemiluminescence must also advance. For instance, the light emission of conventional enhanced luminol substrates substantially decreases within 30 minutes. In contrast, the chemiluminescence signal of the Lumi-LightPLUS substrate persists for more than nine hours, and since exposures of only a few minutes are necessary, multiple exposures can be taken. Additionally, the ability to take extended exposures allows the detection of rare proteins, or higher dilutions of antigen or antibody to be used. The emission wavelength of the Lumi-LightPLUS substrate is 425 nm. Comparison of Lumi-Light Substrate and Lumi-Light PLUS Substrate Lumi-Light Substrate Lumi-LightPLUS Substrate Sensitivity 10-50 pg antigen 1-5 pg antigen Pimary Antibody Dilution Dilute up to 10-fold more compared to colorimetric detection Dilute up to 100-fold more compared to colorimetric detection Signal Stability >3 hours (enhance signal with a 30-minute incubation) > 9 hours (enhance signal with a 30-minute incubation) Number of tests: The reagent is sufficient for 10 blots, each with a size of 10 cm x 10 cm. Sensitivity: Depending on the affinity of the primary antibody, 1 - 5 pg of antigen can be detected in research studies. Storage/stability: The Lumi-LightPLUS substrate components are stable at +2 to +8°C until the expiration date printed on the label. Components are shipped at +15 to +25°C. |
| Other Notes: | For life science research only. Not for use in diagnostic procedures. |
| Packaging: | 1 kit containing 2 components |
| Preparation Note: | Stabilizers: proteinaceous stabilizers Working concentration: The working concentration of conjugate depends on application and substrate. The following concentrations should be taken as a guideline: • Dot blot: 50 mU/ml • ELISA: 25 mU/ml • Western blot: 50 mU/ml Working solution: Preparation of Additional Reagents and Solutions The table inside the package insert describes the preparation of working solutions. Volumes are designed for a membrane of 10x10 cm. If larger membranes are used, volumes have to be scaled up. For reproducible results, equilibrate all solutions to room temperature before use. Note: Do not use Azide to stabilize the solutions against microbial growth, as azide irreversibly inhibits horseradish peroxidase. |
| Quality: | Each lot is function tested using mouse or rabbit primary antibodies. |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 1 |
| Flash Point(F) | does not flash |
| Flash Point(C) | does not flash |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352204 |
| Components | Lumi-LightPLUS Luminol/Enhancer Solution; Lumi-LightPLUS Stable Peroxide Solution |