Enhanced Diagnostic Tools
Non-radioactive Horseradish Peroxidase (HRP) oligonucleotide probes are highly sensitive diagnostic tools. HRP DNA probes have been used for in situ hybridization methods for the detection, enumeration, and localization of specific target sequences in whole fixed cells. Horseradish peroxidase is a 45 kDa large enzyme isolated from horseradish plants. It catalyzes the oxidation of many substrates in the presence of H2O2, by converting non-fluorescent reduced form to fluorescent oxidized forms. Studies have shown that by using a combination of enzyme HRP probes and the tyramide signal amplification system (TSA), also known as CARD-FISH, FISH signals are up to 20-fold brighter. (Schönhuber et al., 1997). This "Catalyzed Reporter Deposition - Fluorescent in situ hybridization" (CARD-FISH) method represents an excellent tool for quantitative detection of microorganisms.
Bio-Synthesis’ HRP FISH probes are manufactured under strict quality control using mass spectrometry, analytical HPLC and HRP enzymatic activity assay.
Price: Standard price covers the labor and materials, including the cost of HRP and it's labeling, cost of maleimide-activated oligo preparation, conjugation procedure, conjugate purification, and analysis. Please contact us for a quotation.
Discount: A 15% discount price applies to additional conjugates ordered at the same time.
Our Oligonucleotide HRP labeling is based on one step conjugation of 5'-modified oligonucleotide directly linked to several available HRP carboxylates via EDC coupling method. This method is superior to the use of a two-step process as provided by other vendors. There is no need to introduce bifunctional cross linker. This method results in greater than 90% coupling efficiency that affords a desired 1:1 oligonucleotide/protein conjugates. Final conjugate are further purified by HPLC chromatography. PAGE is not recommended as it impairs protein activity.
The above image demonstrates oligo conjugation purity and functional assay, showing the activity of enzyme after conjugation.