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Encyclopedia > Secretion assay

Secretion assay is a process used in cell biology to identify cells that are secreting a particular protein (usually a cytokine). It was first developed by Manz et al in 1995. Cell biology (also called cellular biology or cytology, from the Greek kytos, container) is an academic discipline which studies cells. ... Secretion is the process of segregating, elaborating, and releasing chemicals from a cell, or a secreted chemical substance or amount of substance. ... A representation of the 3D structure of myoglobin, showing coloured alpha helices. ... Cytokines are small protein molecules that are the core of communication between immune system cells, and even between these cells and cells belonging to other tissue types. ... 1995 (MCMXCV) was a common year starting on Sunday of the Gregorian calendar. ...


Usually, a cell that is secreting the protein of interest is isolated using an antibody-antibody complex that coats the cell and is able to "catch" the secreted molecules. The cell is then detected by another fluorochrome-labelled antibody, and is subsequently extracted using a process called fluorescent-activated cell sorting (FACS). The FACS method is broadly similar to the enzyme-linked immunosorbent assay (ELISA) antibody format, except that the encapsulated cells remain intact. This is advantageous as the cells are still living after the extraction has taken place. A fluorophore is a fluorescent molecule. ... Schematic of antibody binding to an antigen An antibody is a protein used by the immune system to identify and neutralize foreign objects like bacteria and viruses. ... Fluorescent-activated cell sorting is a type of flow cytometry, a method for sorting a suspension of biologic cells into two or more containers, one cell at a time, based upon specific light scattering and fluorescent characteristics of each cell. ... The Enzyme-Linked Immunosorbent Assay (ELISA or EIA for short) is a biochemical technique used in immunology to detect the presence of an antibody or an antigen in a sample. ...


Further advances now mean that it is possible to extract the secreting cells using a magnetic-based seperation system or in a flow cytometer. Flow cytometry is a technique for counting, examining and sorting microscopic particles suspended in a stream of fluid. ...


A number of commercial applications exist for secretion essay. One such example is the Gel Microdrop (GMD) techonology, developed by One Cell Systems. One Cell asserts that GMD typically recovers a higher number of viable secreting cells than other methods, whilst ignoring any cells which are not secreting the desired protein.


External links

  • One Cell Systems - Gel Microdrop (GMD) Secretion Assay techonology.
  • Nature article - Cell identification and isolation on the basis of cytokine secretion: A novel tool for investigating immune responses.

References

1. Manz et al. (1995) Proceedings of the National Academy of Sciences USA The Proceedings of the National Academy of Sciences of the United States of America, usually referred to as PNAS, is the official journal of the United States National Academy of Sciences. ...



Proteins: key methods of study

Protein methods | Protein purification | Protein structure prediction | Green fluorescent protein | Western blot | Protein immunostaining | Protein sequencing | Gel electrophoresis | Protein immunoprecipitation | Protein structural alignment | Protein ontology | Peptide mass fingerprinting A representation of the 3D structure of myoglobin, showing coloured alpha helices. ... Protein methods are the techniques used to study proteins. ... Protein methods are the techniques used to study proteins. ... Protein purification is the process of isolating proteins from a homogenate, which may comprise cell and tissue components, including DNA, cell membrane and other proteins. ... Protein structure prediction is one of the most significant tasks tackled in computational structural biology and theoretical chemistry. ... GFP ribbon diagram from PDB database The green fluorescent protein (GFP) is a protein from the jellyfish Aequorea victoria that fluoresces green when exposed to blue light. ... Picture of a Western blot with 5 vertical lanes A western blot (a. ... Immunostaining is a general term in biochemistry in that applies to any use of an antibody and some colouring agent to detect a specific protein in a sample. ... Proteins are found in every cell and are essential to every biological process, protein structure is very complex: determining a proteins structure involves first protein sequencing - determining the amino acid sequences of its constituent peptides; and also determining what conformation it adopts and whether it is complexed with any... SDS-PAGE autoradiography DNA agarose gel Gel electrophoresis is a group of techniques used by scientists to separate molecules based on physical characteristics such as size, shape, or isoelectric point. ... Immunoprecipitation is the technique of precipitating an antigen out of solution using an antibody specific to that antigen. ... Protein structural alignment is a form of alignment which tries to establish equivalences between two or more protein structures based on their fold. ... Protein ontology or Proteome Ontology is a research tool of proteomics, similar to the scientific classification system used in biology. ... Peptide mass fingerprinting (PMF) is an analytical technique for protein identification that was developed by John Yates and colleagues (3). ...


Assay: Enzyme assay | Protein assay | Secretion assay An assay is a procedure where the concentration of a component part of a mixture is determined. ... Enzyme assays are laboratory methods for measuring enzymatic activity. ... The Bradford protein assay is a spectroscopic analytical procedure used to measure the concentration of protein in a solution. ...


 
 

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