Common absorbance applications and assays including ELISAs, nucleic acid and protein quantitation, and microbial growth.

AlphaScreen

AlphaScreen® Technology is a homogeneous, high-throughput method for detecting intramolecular binding. Donor and acceptor beads are attached to biomolecules, and when the beads are brought into proximity (via binding), a cascade of chemical reactions occurs, amplifying a signal that is detected on the SpectraMax® i3x and Paradigm® Multi-Mode Microplate Readers.

Cell Health

Cell viability refers to the number of healthy cells in a population and can be evaluated using assays that measure enzyme activity, cell membrane integrity, ATP production, and other indicators. These methods can employ luminescent, fluorescent, or colorimetric readouts as indicators of general cell viability or even specific cellular pathways. Cytotoxicity and cell viability assays are often used to assess a drug or other treatment’s effect, and are valuable tools in the search for new therapeutics, as well as advancing our understanding of how normal cells function.

Cellular Signaling

Cellular signaling allows cells to respond to their environment as well as to communicate with other cells. Proteins located on the cell surface can receive signals from the surroundings and transmit information into the cell via a series of protein interactions and biochemical reactions that comprise a signaling pathway. Multicellular organisms rely upon an extensive array of signaling pathways to coordinate the proper growth, regulation, and functioning of cells and tissues. If signaling between or within cells is dysregulated, inappropriate cellular responses may lead to cancer and other diseases.

Drug Discovery & Development

For every drug that makes it to the finish line, another nine don’t succeed. This alarming failure rate can be traced to reliance on 2D cell cultures that don’t closely mimic complex human biology, often leading to inaccurate predictions of a drug’s potential and extended drug development timelines.

ELISA

Enzyme-linked immunosorbent assays (ELISAs) are used to measure the amount of a specific protein, using a microplate format, and results are most often detected via absorbance in the visible wavelength range. Chemiluminescent and fluorescent ELISA formats offer enhanced sensitivity for accurate quantitation of less abundant analytes.

Fluorescence

Many fluorescence-based assays include cell viability, GPCR activity, and fluorescent nucleic acid quantification.

Fluorescence Polarization (FP)

Fluorescence polarization (FP) is a technique that is widely used to monitor binding events in solution. It can be used to assess biomolecular interactions, including protein-antibody binding and DNA hybridization, as well as enzyme activity, and it has been adapted to basic research as well as high-throughput screening.

IgG Quantification

In therapeutic protein engineering and cell line development, IgG quantification is the process of measuring the amount of immunoglobulin G (IgG), a common class of therapeutic proteins, produced by a genetically modified cell line. This is important for evaluating and monitoring the productivity of the cell line and selecting the best candidate for further development of therapeutic antibodies.

Kinase/Phosphatase Assays

Kinases are one of the most critical targets in drug discovery today. These enzymes are key components in cellular signaling pathways, and disruption to their function causes a variety of diseases, such as metabolic diseases, certain cancers, and cardiac disease. The functionally related phosphatases and phosphodiesterases are also important screening targets.

Luminescence

The key luminescence-based assays include dual-luciferase reporter gene, chemiluminescent ELISA, cytotoxicity, and BRET.

Microbiology and Contaminant

Microbes, including bacteria, have been estimated to make up about 15 percent of the earth’s biomass, and microbes in the human body outnumber human cells by 10 to 1. These microorganisms provide great benefit to us and are also vital to many fields of research from medicine to alternative energy production. On the other hand, monitoring for microbes and the toxic substances they produce is necessary to ensure the safety of pharmaceutical products. Scientists whose research relies on mammalian cells must carefully monitor these cultures for unwanted microbial contaminants to ensure that their experimental results are reliable.

Nanoparticles

Nanomaterials are typically less than 100 nm in diameter, small enough to penetrate mammalian cells. They can be synthesized in many shapes, such as rods, tubes, and particles, as well as in different materials. Targeting or therapeutic molecules can be readily attached to nanoparticles, and when these are delivered systemically, the targeting molecules enable detection of certain cell populations, such as tumor cells, while attached therapeutic compounds can act on the targeted cells.

Nucleic Acid (DNA/RNA) Quantitation and Analysis

Nucleic acids are large biomolecules common to all known life forms. Deoxyribonucleic acid (DNA) consists of a double strand of pairs of nucleotides, while ribonucleic acid (RNA) is typically a single strand. In DNA, the nucleotides are adenine, cytosine, guanine, and thymine, while RNA contains uracil instead of thymine. DNA makes up the genetic material of all organisms, encoding the information cells need to synthesize proteins.

Protein Detection, Quantitation and Analysis

Protein detection, quantitation, and analysis are central to the investigation of a wide variety of biological processes. Measuring the concentration of protein is necessary to processes ranging from protein purification and labeling to sample preparation for electrophoresis. Protein can be quantitated directly via absorbance at 280 nm, or indirectly using colorimetric (BCA, Bradford, etc.) or fluorometric methods offering advantages such as greater sensitivity. To identify and measure a specific protein within a complex sample, for example, serum or cell lysate, an ELISA may be used.

Spheroids in 3D biology

Cancer is a leading cause of death worldwide, but researchers are making enormous strides towards understanding its underlying mechanisms and working towards new treatments. Spheroids, 3D aggregates of cells in culture, have emerged as a valuable research tool that provides more physiological relevance than traditional 2D cell culture

TRF, TR-FRET và HTRF

Time-resolved fluorescence – TRF and TR-FRET including HTRF assays provide applications resources to aid in your research, including kinase assays, cellular signaling pathways, protein-protein interactions, cell cytotoxicity, and more.

Western blotting

Western blotting is a popular technique used for protein detection and quantitation. Molecular Devices introduce a unique Western blot detection system on a multi-mode microplate reader, including colorimetric, fluorescence, and chemiluminescence detection methods.