The SAM2® Biotin Capture Membrane binds biotinylated molecules based on their affinity for streptavidin. The proprietary process by which the SAM2® Membrane is produced results in a high density of streptavidin on the filter, providing rapid, quantitative substrate binding in the nmol/cm2 range, depending upon the substrate used. In addition, the membrane is designed to minimize nonspecific binding. The membrane is available either as a large, prenumbered, partially cut sheet (approximately 10.5 × 15.0cm; Cat.# V2861) or as a smaller, uncut sheet (approximately 7.6 × 10.9cm; Cat.# V7861). The partially cut membrane allows easy separation into 96 individual squares and is designed for small-scale experiments where high binding capacity is required. The uncut sheet can be analyzed as a whole membrane or may be cut to the size desired. The uncut membrane allows for sample application using a multichannel pipettor. Both membranes may be analyzed using phosphorimaging analysis, autoradiography or scintillation counting to quantitate results. The membranes have also been used successfully with chemiluminescence detection techniques. The use of fluorescence for detection of captured molecules is not recommended at this time.
Features - Benefits
Use a Variety of Substrates: Analysis of biotinylated substrates can be applied to a wide variety of substrate types without the need to optimize each substrate for binding to a matrix. The user can perform experiments with a wide array of sample numbers and sizes without changing the analysis technique, since the membrane is available in 96-square (partially cut) and solid sheet (uncut) formats. Minimize Nonspecific Binding: The combination of protein denaturant and high-salt washes minimizes nonspecific binding to the membrane without interfering with the high-affinity interaction between streptavidin and biotin. Obtain High Signal-to-Noise Ratios: The stringent washing conditions employed assist in attaining very low background counts. Perform Kinetic Studies: Membrane can linearly bind biotinylated substrates up to the nmol/cm2 range. Allows for kinetic studies. Strong Binding Reaction: Membrane retains the biotin conjugate over 8 logs of pH (pH 2–10), changes in temperature, organic solvents, ionic and nonionic detergents (SDS, CHAPS, Triton® X-100, Tween® 20 and Tween® 80) and denaturing agents (5M guanidine-HCl and 2M urea). Rapid: Binds within 1 minute. Convenient: Compatible with enzyme assays using radioactive detection. Membranes manufactured by this method have been shown to allow chemiluminescent detection.
Applications
Protein kinase activity quantitation.
Specific mRNA quantitation.
Cytokine quantitation.
Specific gene quantitation.
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