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Streptavidin is widely used in Western blotting and immunoassays conjugated to some reporter molecule, such as horseradish peroxidase. Streptavidin has also been used in the developing field of Nanobiotechnology, the use of biological molecules such as proteins or lipids to create nanoscale devices/structures.
Here, the antibody provides the specificity to locate the protein of interest, and the HRP enzyme, in the presence of a substrate, produces a detectable signal. [7] Horseradish peroxidase is also commonly used in techniques such as ELISA and Immunohistochemistry due to its monomeric nature and the ease with which it produces coloured products ...
Also, the Strep-tag system can be used for protein detection in various assays. Depending on the experimental circumstances, Strep-tag antibodies or Strep-Tactin, with an enzymatic (e.g.horseradish peroxidase (HRP), alkaline phosphatase (AP)) or fluorescence (e.g. green fluorescent protein (GFP)) marker. If high purity is required, the lysate ...
The primary antibody can be tagged with a small molecule that can be recognised by a peroxidase-conjugated binding molecule with high affinity. The most common example of this is a biotin linked primary antibody that binds to an enzyme-bound streptavidin. This method can be used to amplify the signal. Example 3.
Horseradish peroxidase is commonly linked to secondary antibodies to allow the detection of the target protein by chemiluminescence. The chemiluminescent substrate is cleaved by horseradish peroxidase, resulting in the production of luminescence. Therefore, the production of luminescence is proportional to the amount of horseradish peroxidase ...
This tag can also be used in detection of the protein via anti-biotin antibodies or avidin/streptavidin-tagged detection strategies such as enzyme reporters (e.g., horseradish peroxidase, alkaline phosphatase) or fluorescent probes.
The Streptavidin-Binding Peptide (SBP)-Tag is a 38-amino acid sequence that may be engineered into recombinant proteins. Recombinant proteins containing the SBP-Tag bind to streptavidin and this property may be utilized in specific purification, detection or immobilization strategies.
When using a biotin label, streptavidin conjugated to an enzyme such as horseradish peroxidase is used to detect the DNA fragment. [ 8 ] [ 9 ] While isotopic DNA labeling has little or no effect on protein binding affinity, use of non-isotopic labels including flurophores or biotin can alter the affinity and/or stoichiometry of the protein ...
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