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Alkaline phosphatase: (1) a phosphatase enzyme found in high concentrations in bone, liver, kidneys, and placenta, is active in an alkaline medium. (2) an enzyme covalently attached to an antigen in a competitive immunoassay
Antibody: a protein produced by B cell lymphocytes in response to a particular antigen; an immunoglobulin that recognizes some portion of an antigen molecule
Analyte: a molecule that is measured or analyzed
Antigen: a substance that evokes an immune response and is specifically bound by a given antibody
AP: see alkaline phosphatase
Apoptosis: a process of programmed cellular selfdestruction
Aprotinin: a naturally occurring biochemical that inhibits plasmin, as well as trypsin and chymotrypsin enzyme activities
B0: a measurement of the binding maximum in competitive immunoassays; in an immunoassay, the B0 well typically contains antibody, enzyme-linked conjugate, and buffer.
Bioluminescence: a luminescent process mediated by an enzyme or other biological system that causes an organism to glow
Biomarker: a substance or molecule used as an indication of the presence or absence of a certain disease state
Buffer: a system capable of resisting changes in pH when small quantities of acid or base are added
Chemiluminescence: the release of light as the result of a chemical reaction
Chemiluminescent immunoassay: a type of immunoassay in which the enzyme conversion of substrate results in the emission of light, detectable at a specific wavelength and measured in Relative Light Units
Chemokine: a cytokine that stimulates chemotaxis in white blood cells
CLIA: see chemiluminescent immunoassay
Coefficient of variation: a statistical expression of precision based on the standard deviation and average of multiple measurements; %Coefficient of Variation = (Standard Deviation/Mean) * 100
Coelenterazine: a chromophore that is the substrate for several marine bioluminescent reactions or which is incorporated as part of a photoprotein; often used to reconstitute aequorin in cells transfected with apoaequorin cDNA
Competitive immunoassay: an assay format used to detect small molecules; involves the use of a specific antibody which binds the antigen in the sample along with the antigen which has been labeled with a detection molecule such as alkaline phosphatase; the amount of bound conjugate is inversely proportional to the amount of unlabeled antigen in the sample.
Conditioned media: cell culture media that has been exposed to cells; includes all added supplements in addition to the molecules secreted by the cells themselves.
Conjugate: two covalently-linked molecules; an antibody or antigen that has been labeled with an enzyme tracer
Cross reactivity: a measurement of how efficiently an antibody can measure molecules other than the antigen it was
raised against
CV: See coefficient of variation
Cyclic nucleotide: a derivative of nucleic acid that regulates a number of metabolic activities; often referred to as a “second messenger”; activated by hormones and neurotransmitters.
Cytokine: a secreted protein that binds to a specific cell surface receptor and initiates a cascade of events affecting a variety of cellular functions
Diluent: buffer or liquid medium used to dilute a standard or sample
Drift: the difference in signal seen from one side of a microtiter plate to the other
Dynamic range: the continuous span of high to low analyte concentrations that can be reliably detected in an immunoassay
ED50: (1) a common measurement of sensitivity in a competitive immunoassay; analyte concentration when the %B/B0 is at fifty percent; (2) a measurement of the dose of a drug that is pharmacologically effective for fifty percent of the population or a fifty percent response in a biological system exposed to the drug
Edge effect: the difference in signal between the exterior and interior wells of an immunoassay’s microtiter plate
EIA: see enzyme immunoassay
Eicosanoid: a class of C20 arachidonic acid derivatives that include prostaglandins, thromboxanes, and leukotrienes
ELISA: see enzyme linked immunosorbent assay
Enzyme: a type of protein that acts as a catalyst
Extraction efficiency: a calculation used to determine the true analyte concentration when extraction protocols are necessary for measurement; the amount of recovered analyte is divided by the amount of analyte in the pre-extraction sample and expressed as a percentage
Flow Cytometry: a method for measuring various components or structural features in cells; involves the use of a laser beam to detect the light-absorbing or fluorescing components of cells.
Fluorescence: light emitted by excited molecules as they revert to their ground state
Fluorescence polarization immunoassay: An immunoassay that uses measurement of polarized light (milli-polarization unit, or mP) as a readout of antigen concentration. Antigen conjugated to a fluorescent tracer (ie FITC) competes with antigen in the sample for binding to an antibody in solution. When excited by polarized light, tracer bound by antibody tumbles in solution at a slower rate than unbound tracer, resulting in an increase in polarized light and larger mP. In this way, the readout is similar to a competitive EIA format, where readout (mP) is inversely proportional to the amount of antigen in the sample.
FPIA: see fluorescence polarization immunoassay
Heat shock protein: any of a class of molecular chaperone proteins synthesized in response to environmental stressors such as extremes in temperature
Hormone: a biochemical substance synthesized by one type of cell or tissue that produces a specific effect on a second type of cell or tissue
Horseradish peroxidase: an enzyme commonly used as a label for antibodies
HRP: see horseradish peroxidase
HSP: see heat shock protein
Hybridoma: a cell line formed from the fusion of the splenocytes of an immunized animal and myeloma cells
ICC: see immunocytochemistry
IHC: see immunohistochemistry
Immunocytochemistry: an antigen detection technique using an antibody to detect antigen in cultured cells
Immunoglobulin: any of a group of large glycoproteins, composed of two heavy and two light chains, that function as antibodies in the immune response and are secreted by plasma cells
Immunohistochemistry: an antigen detection technique using a fluorophore- or enzyme-tagged antibody to detect an antigen in tissue
Immunometric immunoassay: an assay format used to detect large molecules; involves a solid phase coated with an antigen-specific antibody which binds the antigen in the samples; a secondary antibody labeled with an enzyme is then added and detected through the use of a substrate.
Interference: a condition that prevents the completion of an unrestricted competitive binding reaction and its subsequent detection in an immunoassay; this can be caused by the presence of certain antibodies (species interference), an overwhelming amount of sample constituents (matrix effect) or inappropriate chemicals in a sample.
Interferon: a member of a family of small proteins that stimulates viral resistance in cells
Isotypes: the classes of immunoglobulins which differ in molecular weight and concentration in the blood: IgG, IgA, IgM, IgD, and IgE
Leukotriene: an arachidonic acid derivative that is an important mediator of inflammation and allergic reactions
Linearity: the ability to consistently detect the same amount of antigen through multiple serial dilutions
Lysate: The contents released from a lysed cell
Matrix: the environment in which something is found; used to refer to the solid matrix that is used during an assay (e.g., tubes or microtiter wells) or the source of the sample (e.g., serum, plasma, saliva, urine, media, etc).
Matrix effect: a type of interference caused by a constituent of the sample itself; usually relates to the pH, osmolarity or composition of the sample used in an immunoassay; if the sample characteristics exceed the limitations tolerated by the assay, a matrix effect will result and sample detection becomes non-linear.
Monoclonal antibody: a type of antibody derived from hybridoma cells which are of higher purity and specificity than polyclonal antibodies
Neat sample: an undiluted or unaltered sample
Non-conditioned media: culture media containing all supplements required for growth, but which has not been exposed to cells; used as standard diluent when samples are conditioned media
Non-specific binding: a measurement of enzyme activity not caused by antigen-antibody binding in an immunoassay
NSB: see non-specific binding
OD: see optical density
Optical density: an expression of absorbance for an optical element at a given wavelength
Oxidative stress: a condition that occurs resulting from an excess of free radicals, a decrease in antioxidant levels, or both
Polyclonal antibody: an antibody that is produced by more than one type of B-cell
Precision: a statistical evaluation of the ability to detect the same value over multiple measurements.
Prostaglandin: a C20 unsaturated fatty acid which is a metabolite of arachidonic acid; a local autacoid that forms rapidly and regulates cellular activities by exerting its effects locally
Sample recovery: a statistical expression of the ability to measure antigen that has been added, or spiked, into the sample; the dilution factor required for an approximate 100% recovery of the added antigen is the recommended minimum dilution to avoid a matrix effect.
Sandwich immunoassay: see immunometric immunoassay
Serial dilution: a set of successive dilutions where the prior dilution step serves as the sample source for the next dilution step
Sensitivity: a measurement of the smallest increment that can be reliably detected in an immunoassay; the calculated value based on optical density statistical data from the B0 and lowest concentration standard
Spiked sample: a sample into which a known concentration of analyte has been added
TA: see total activity
Total activity: a measurement of the maximum enzymatic activity expected for an immunoassay; wells will typically contain a specific amount of the detection enzyme molecule and substrate
Western blotting: a technique for identifying a particular protein using antibodies after eletrophoretic separation in a gel and transfer to a membrane