Lumitera is a One-Component Chemiluminescent Membrane / Blotting Peroxidase Substrate is luminol based and can detect horseradish peroxidase at high sensitivity levels (low picogram to femtogram). It provides superior sensitivity and convenience compared to competitor products. The substrate is supplied as ONE component. Lumitera may be used for any blotting application utilizing horseradish peroxidase (HRP)-conjugates. The substrate can be used with various blocking buffers and on nitrocellulose or PVDF membranes. Such blots will exhibit low backgrounds. Detection and analysis may be done by CCD imaging systems or x-ray film. Lumitera is a one component ready to use reagent providing unique convenience, sensitivity and reduced costs for kit manufacturers.
◊ No mixing is required.
◊ Get consistent results by avoiding aliquoting and mixing errors.
◊ Kit manufacturers can reduce costs by eliminating additional bottles and superfluous packaging.
Please perform western blotting for the primary and secondary antibody reactions in a general way.
1. Place Lumitera at room temperature 1-2 hours before usage.
2. Eliminate excess buffer from the washed membrane. Place the membrane on top of plastic wrap with the protein side facing upwards. Add 100ul of Lumitera per 1cm2 of membrane. Leave at room temperature.
3. Chmiluminescence should be able to to be observed 1 to 10 minutes after adding Lumitera.
◊ Equilibrate an aliquot of Lumitera at room temperature before use. Aliquot into a clean container. Do not contaminate the substrate with HRP enzyme or other proteins. Never pipette directly from the Lumitera stock substrate storage bottle or pour used or aliquoted solution back into the stock vessel.
◊ Avoid increasing backgrounds by handling Blots with clean gloves and clean forceps. Forceps contaminated with rust can lead to an unwanted reaction and increased backgrounds.
◊ Analytes can be applied to membranes as a dot blot or via gel transfer. Blotting conditions should be optimized for each assay system. Use approximately 100 uL of Lumitera per square centimeter of membrane.
◊ Place membrane in a clean, dry vessel. Add Lumitera to the membrane and incubate at room temperature for optimal detection. Best results for chemiluminescence can be obtained from one to 10 minutes after contacting substrate with HRP enzyme.
◊ Remove excess substrate by blotting on filter paper. Cover membrane with clear plastic wrap and visualize by either x-ray film or a CCD imaging system.
◊ Lumitera substrate has a wide range to detect HRP enzyme on a membrane.
|Figure 1. A comparison of Lumitera with Company A low-sensitivity and high sensitivity blotting substrates.||Figure 2. A comparison of Lumitera with Company B and C. Lumitera exhibits comparable data to Company B high sensitivity blotting substrate.|
To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.