Antibody Techniques and Codes

Antibodies can be used for a wide variety of techniques and purposes. It is important to understand what techniques your antibody is and isn’t compatible with. For example, an antibody which recognises a conformational epitope may be appropriate for ELISA but not for western blots. Our products come with application codes which show which applications each antibody has been used for in the literature. The absence of an application code does not necessarily mean that the antibody is not compatible with that application, only that it has not been tested for that purpose yet.

CategoryTechniqueCodeUse
Detection/QuantificationEnzyme-linked immunosorbant assayELISAMany purposes including subtype determination and specificity determination. There are multiple types of ELISA, with direct ELISA, sandwich ELISA and competition ELISA the most used in literature.
Detection/QuantificationFluorescence-Detection Size-Exclusion ChromatographyFSECAnalysis of SEC coupled with GFP fluorescence analysis to determine oligomeric states. Can be used as a screening method in crystallography.
Detection/QuantificationLateral Flow AssayLFALFAs using antibodies are also known as LFIAs. This is a paper-based assay used for rapid diagnostic of analytes. Often used for medical diagnostics.
Detection/QuantificationPlaque Reduction Neutralisation Testneutralisation assayUsed to quantify the titer of neutralizing antibody for a virus. This code includes use in microneutralization assays.
Detection/QuantificationRadioimmunoassayRIARIA is a sensitive method for detecting the presence of an antigen. It has been developed to measure up to picomolar concentrations of some antigens. A radioactive form of the antigen is used, and then the antibody is added. The bound and free antigen is then separated and the radioactivity of either the bound or unbound fraction is measured and can be converted into a concentration via a standard curve.
Detection/QuantificationRadiolabellingradiolabellingA catch-all code for methods of labelling antigen targets using radiolabelled antibodies. This is used as a catch-all code for radioactive antibody studies (other than RIA), for example IRMA. This does not include radioimmunotherapy which is classed under therapeutic.
Detection/QuantificationVirus Quantificationvirus quantificationA catch-all code for any technique used to quantify virus titres other than plaque reduction neutralisation test.
Detection/QuantificationWestern blotWBNormally immunoblot is used interchangeably with western blot, although technically immunoblot is an umbrella term including dot blot and slot blot. These techniques rely on the same priciples so if an antibody has been used for one it will be possible to use if for any of the others. These techniques are semiquantitative and measure the presence of denatured antigens. This means that only antibodies which recognise linear epitopes can be used in western blots.
ImagingCytometric Bead ArrayCBAThis is a flow cytometry application where antibodies are bound to fluorescent beads with different intensities so multiple beads can be run in a single tube.
ImagingElectron MicroscopyEMAntibodies can have a range of uses in all types of EM especially scanning EM. For example, they can be used to deliver gold nanoparticles to specific sites to provide an area of strong contrast for scanning EM imaging.
ImagingFlow cytometryFCAntibodies mark the presence of a particular antigen/epitope by conjugation to a fluorescent or coloured label. The read-out from a large number of individual cells can then be analysed. This is a powerful technique for analysing sub-populations.
ImagingIHC frozenIHC-FA type of IHC where typically the sample is frozen. Antibodies used in this type of IHC can probably but not necessarily be used in IHC-P.
ImagingIHC paraffinIHC-PA type of IHC which typically used paraffin. The antibodies used in this technique can probably but not necessarily be used in IHC-F.
ImagingIHC resinIHC-RA type of IHC which used a resin. The antibodies used in this technique can probably but not necessarily be used in other forms of IHC so if it says which IHC was used put it down.
ImagingImmunocytochemistryICCThis is similar to IHC except individual cells outside of the context of tissue samples are visualised. This is easier to do but further from physiological conditions. For example used to visualise cells in cell culture.
ImagingImmunofluorescenceIFA catch-all code for techniques which use fluorescently labelled antibody to bind to a protein (except IHC and ICC). For example indirect immunofluorescence tests or SDS-FRL.
ImagingImmunohistochemistryIHCThe process of selectively imaging antigens in cells in a section of biological tissue (contrast to ICC). The antibody is conjugated to a labelling molecule to allow visualisation. To allow the antibodies to permeate the cells they are preserved so they keep their architecture and then ‘fixed’ to allow permeation.
ImagingX-ray crystallographycrystallographyAntibodies are often used to facilitate process of crystallisation, to increase protein solubility and aid crystal packing.
Protein ManipulationAffinity ChromatographychromatographyAntibodies are often used on chromatography columns in an affinity purification step. This can also be used to deplete a sample of a particular antigen by using the flowthrough.
Protein ManipulationDepletiondepleteWhere antibodies are used to remove a particular antigen from a mixture. This is used in the process of negative selection.
Protein ManipulationEnrichmentenrichWhere antibodies are used to enrich a mixture for its target. An example is when this is coupled to MRM resulting in immunoMRM.
Protein ManipulationEpitope Tag Bindingepitope tagAntibodies used to recognise short sequence motifs (epitope tags) fused to proteins of interest. Often used for purification and detection of target proteins.
Protein ManipulationImmunoprecipitationIPUsed to bind and separate proteins bound to the antibody. Co-IP experiments pull down proteins bound to the antigen. Agarose beads are bound to protein A or protein G, which are bacterial proteins which effeciently bind IgG antibodies. The mixture is then spun and the beads pellet pulling down the target protein (and bound proteins) with it.
Binding AssaysBio-layer interferenceBLIAnalyses binding by measuring changes in a visible light interference pattern.
Binding AssaysIsothermal Titration CalorimetryITCMeasures thermodynamic properties of interactions of binding, such as binding affinity, enthalpy changes and Gibbs energy changes. This is achieved by titrating in the ligand and measuring the temperature change by comparison with a reference cell, and keeping the cells at the same temperature. Any difference in energy used to maintain the chambers at the same temperature must be due to the interaction, and this can be used to determine the thermodynamic parameters.
Binding AssaysMicroscale thermophoresisMSTA binding assay which is based on detection of temperature-induced changes in fluorescence. It can be used to determine affinities of molecules and in competition assays.
Binding AssaysSurface plasmon resonanceSPRGives dissociation and association constants for antibody binding. Sometimes referred to as ‘Biocore studies’ when using Biocore products for SPR.
Binding AssaysThin layer chromatographyTLCIs sometimes used as way to analyse binding of an antibody to its antigen.
ApplicationDiagnosticdiagnosticThis code is used when the antibody has been has been tested with the aim of developing it into a diagnostic. It is not be used if the antibody has just been suggested to possibly have some diagnostic use without any testing.
Applicationin vivoin vivoWhen the antibody has been used in vivo.
ApplicationNegative Controlnegative controlWhen antibodies are used as a negative control in experiments.
ApplicationPositive Controlpositive controlWhen antibodies are used as positive controls in experiments.
ApplicationTherapeutictherapeuticWhen the antibody has been tested in a therapeutic context (including animal trials). This code is not used if the antibody has just been suggested to possibly have some therapeutic use without any testing.
ApplicationTransgenic StudiestransgenicWhen the antibodies are constitutively expressed in a different species, especially plants or other atypical kingdoms.
ApplicationVaccinationvaccinationSome antibodies have been used in making vaccines, for example targeting the protein to immune cells to elicit a stronger response.
FormatBispecificbispecificWhen an antibody binding domain has been used sucessfully in a bispecific antibody.
FormatHumanisationhumanisationReduces immunogenic response to non-human antibodies in humans.
FunctionalActivationactivateWhen the antibody is used to activate a species, for example by acting as an agonist to a receptor.
FunctionalAntibody-dependent cell-mediated cytotoxicity assayADCCDetermines the cytotoxicity of a particular antibody. Often used to analyse the effectiveness of a particular immunotherapy against self-cells.
FunctionalBactericidalbactericidalKills bacteria.
FunctionalBlockingblockThis is where the antibody is used to prevent the binding of another species to the target antigen. For example blocking binding of a ligand to its receptor.
FunctionalCatalytic AntibodycatalyticAntibody has catalytic activity for any reaction.
FunctionalComplement-dependent cytotoxicityCDCDetermines cytotoxicity dependent on the complement system. Often used to analyse the effectiveness of a particular immunotherapy against self-cells.
FunctionalImmunotoxinimmunotoxinUsed to specifically target and kill cells, often by fusing the antibody to a toxic chemical.
FunctionalNeutralisationneutraliseThis is where the antibody is used to inactivate the function/activity of another species. For example neutralisation of toxin/enzyme activity.
FunctionalSpecific Function AssayfunctionalThis is a catch-all code to describe specific function assays which are unique to an antibody. For example sperm agglutination assays, hemadsorption rosetting assays, and cynomolgus bone marrow CFUe assays are examples of specific function assays which may only be relevant to a handful of antibodies.
DNA HandlingChromatin ImmunoPrecipitationChIPUsed to identify binding of proteins to DNA, by coprecipitation of particular DNA sequences with antigens. This includes ChIP-seq.
DNA HandlingElectrophoretic mobility shift assayEMSAUsed to measure protein binding to DNA and observing a shift in the mobility of the DNA fragment using labelled probes.