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Various labeling techniques such as enzymatic labeling, protein labeling, and genetic labeling are widely utilized. Ethidium bromide, fluorescein and green fluorescent protein are common tags. The most commonly labelled molecules are antibodies, proteins, amino acids and peptides which are then used as specific probes for detection of a ...
Maleimide-mediated methodologies are among the most used in bioconjugation. [5] [6] Due to fast reactions and high selectivity towards cysteine residues in proteins, a large variety of maleimide heterobifunctional reagents are used for the preparation of targeted therapeutics, assemblies for studying proteins in their biological context, protein-based microarrays, or proteins immobilisation. [7]
Fluorescein diacetate (FDA) hydrolysis assays can be used to measure the enzyme activity of microbes in a sample. A bright yellow-green glow is produced and is strongest when enzymatic activity is greatest. This can be quantified using a spectrofluorometer or a spectrophotometer.
Detailed protocols are now available that can be used to label lymphocytes (and other cell types) with a high degree of reliability and precision. [ 22 ] [ 23 ] [ 24 ] One of the most important parameters, however, is to ensure that the cell population being studied has not been too heavily labelled with CFSE, as such cells, although remaining ...
Different indirect methods can be employed to achieve high degrees of specificity and sensitivity. First, two-step protocols are often used to avoid the cross-reaction between the immunolabeling of multiple primary and secondary antibody mixtures, where secondary antibodies Fab fragments are frequently used.
I.e., FluoProbes488 reduces the background in Flow Cytometry [5] and in slide microscopy allowing sharper and brighter images. [6] FluoProbes 488, 547H and 647H are found more photostable [ 7 ] that is taken to good account in applications using long illuminations periods (i.e. scanning such as in confocal microscopy ), [ 8 ] or for longer ...
Fluorescein can be introduced to a plant's veins through the roots or a cut stem. The dye is able to be taken up into the plant the same way as water and moves from the roots to the top of the plant due to a transpirational pull. [29] The fluorescein that has been taken up into the plant can be visualized under a fluorescent microscope.
SNAP-tag reaction scheme. SNAP-tag® is a self-labeling protein tag commercially available in various expression vectors. SNAP-tag is a 182 residue polypeptide (19.4 kDa) that can be fused to any protein of interest and further specifically and covalently tagged with a suitable ligand, such as a fluorescent dye.