The CiteAb Antibody Glossary
The CiteAb glossary: a quick reminder for all those terms that can so easily slip your mind!
Quick Links | ||
Antibodies | Host | Suppliers |
Applications | Reactivity | References |
Antibodies
Term | Description |
Antibody | Antibodies along with their membrane bound counterpart, the B cell receptor, are the immunoglobulin utilised by B cells in the recognition and neutralisation of offensive antigen. Employed by the immune system during an immune response, antibodies serve two main functions; to bind specifically to a complimentary antigen and then to recruit other cells and molecules to destroy the bound pathogen. Structurally these functions are separated within the antibody molecule. |
Antibody Structure | Antibodies are Y-shaped globular proteins of ~150kDa in mass. They consist of two identical heavy and two identical light protein chains made up of ~110 amino acid immunoblobulin domains |
Constant Region | Simply put, the constant region is the end of an antibody that things stick to. Depending on the immunoglobulin isotype of the heavy chains, which cells and molecules that bind will vary. |
Variable Region | The variable region contains the antigen binding site: basically it’s the end that sticks to things! |
Hinge Region | The flexible domain linking the trunk of the Y-shaped structure with the two arms that contain the antigen binding site. |
Heavy Chain | Heavy chains consist of one amino-terminal immunoglobulin domain belonging to the variable region and up to four more constant domains, depending on the isotype. |
Light Chain | The light chain consists of two immunoglobulin domains. Each antibody will contain two identical light chains, one in each arm of the y-shaped structure. |
F(ab’)2 | An antibody when digested with the pepsin enzyme will produce two Fab that remain linked by the hinge region. |
Fab Region | Proteolytic digestion of an antibody molecule with the papain enzyme will yield two Fab and one Fc fragment. These identical Fab form the arms of the Y-shaped antibody molecule and contain four immunoglobulin domains: the two variable region domains of the heavy and light chains that form the antigen binding site and two constant domains. |
Antibody Isotype/Class | Antibodies in placental mammals can be divided into five isotypes or classes based on which heavy chains they possess: classes IgM, IgD, IgG, IgE and IgA. |
IgM | Immunoglobulin M is the first antibody produced during the humoral immune response, prior to any class switching. Once secreted it is capable of forming pentamers with other IgM antibodies, making it the largest antibody in circulation. |
IgD | Immunoglobulin D is co-expressed alongside IgM on the surface of mature B cells. |
IgG | Immunoglobulin G is the most abundant serum antibody making up around 75% of circulating Ig. It can be further divided into 4 subclasses; IgG1, IgG2, IgG3 and IgG4. |
IgG4 | IgG4: Is the least abundant subclass of IgG and has the ability to split down the middle of the Y-shape to form two separate heavy chain-light chain fragments. These fragments are then able to rejoin with other separated IgG4 heavy chain-light chain fragments to form bivalent antibodies, with each arm specific to a different antigen. |
IgE | IgE is the least abundant of the 5 circulating antibody isotypes in humans and typically only occurs at 0.05% the concentration of circulating IgG. The reason for this low concentration is the high affinity with which activated eosinophils, basophils and mast cells are able to bind IgE via their ϵ-heavy-chain Fc receptors (FcϵR). Unlike other antibody isotypes which must have bound antigen, mast cell FcϵR are capable of binding IgE that has not yet bound to antigen, effectively sensitising the cells. |
IgA | Immunoglobulin A is the main antibody secreted by mucosal lymphoid tissues and can be found in both monomeric and dimeric forms. |
IgY | The major serum antibody of birds and reptiles, IgY was first described by Klemperer et al. in 1893 but for the better part of the 20th century was known as IgG due to it’s similarity with the mammalian IgG. |
IgW | |
Epitopes, Antigens and Immunogens | Epitopes are the specific region within an antigen that antibodies recognise and bind to. Those antigens that are capable of eliciting an immune response are known as immunogens. Antigens may also be combined with adjuvants in order to increase their immunogenicity. |
Adjuvant | A substance that increases the immunogenicity of the substance with which it is mixed. Common adjuvants include: incomplete/complete Freuds adjuvant, alum (aluminium hydroxide), alum plus and immune stimulatory complexes (ISCOMs). |
Monoclonal | Monoclonal antibodies are the product of a single cloned B lymphocyte plasma cell. |
Polyclonal | Polyclonal antibodies are the product of multiple B cell clones, because of this not all antibody clones will bind the same epitope within an antigen. |
Host | The animal in which an antibody is raised (if polyclonal), or that provides the B cell component of a hybridoma (monoclonal). |
Reactivity | The species from which an immunizing epitope was derived. Antibodies may also be reactive to other species depending on the sequence homology of the epitope. |
Aptamer | Aptamers, from the Latin aptus, to fit and the Greek meros for “a bit of a thing” are a group of affinity reagents used in much the same way as antibodies. Like antibodies, and as you would expect from an affinity reagent they bind with high affinity to specific targets. Unlike antibodies however, aptamers are created in vitro and can come in a number of different formats: nucleic acid (RNA/DNA/XNA) or peptide based. |
Spiegelmer | Spiegelmers, from the German spiegel “mirror”, are RNA aptamers constructed of L-ribose units rather than the D-ribose units which occur more commonly in Nature. Being constructed from L-ribose units makes spiegelmers more resistant to degradation by nucleases. |
Applications
Application | Description |
Affinity Chromatography | Affinity chromatography is a method used to separate out antigen or antibody from a sample based on specific antibody-antigen interaction. A simple columnar chromatography setup involves adding sample to a column containing immobilised antigen (agarose beads are often used). Antibodies specific to the immobilised antigen remain bound in the column while the remainder of the mixture passes through. Following a washing step to remove any remaining unbound sample, an elution buffer is added to elute the bound antibody, which is then collected. |
Control | Antibodies when used as a control are of the same isotype as the primary detecting antibody, but importantly have no relevant specificity to the target antigen. They’re used to help differentiate specific antibody binding from non-specific, background noise. |
Diffusion | Diffusion based assays (of which there are a number of types) involve the diffusion of antigen and antibody through a uniform medium such as agar. |
Dot Blot | A dot blot (also known as slot blot) can be thought of as a simplification of the Western blot, minus the electrophoretic separation of sample prior to detection with antibodies. |
EIA | An Enzyme ImmunoAssay is any one of several techniques that use an enzyme conjugated antibody. EIA’s include; ELISA, cELISA, ELISpot. |
Electrochemiluminescence | |
ELISA | Enzyme Linked ImmunoSorbent Assays are a method for the quantitation of antigens and antibodies. Simply, sample antigens are fixed to a surface such as a microtitre plate and enzyme conjugated antibodies specific to the target antigen are allowed to bind. The enzyme’s substrate is then added to produce a detectable signal, usually a colour change. Alternatively as in sandwich-ELISA, the detecting antibody is bound to a surface and an antigen containing sample added. A second enzyme conjugated detecting antibody can then be added to bind the antigen forming an antibody-antigen-antibody “sandwich”. Once again the enzyme’s substrate is added to produce a detectable signal. |
cELISA | *In cell ELISA |
ELISpot | Enzyme Linked ImmunoSorbent spot assays are a modified ELISA method for the ex-vivo detection of secreted antigens (cytokines/antibodies). Unlike ELISA, which allows determination of antigen concentration, ELISpot allows quantification of the number of cell within a culture that are secreting the antigen of interest. The ELISpot method is very similar to sandwich ELISA but with a culture of secreting cells added to the plate. The antigens released by these cells, for example cytokines, are detected by the primary detection antibodies bound to the plate before they can be reabsorbed by other cells within the culture. As in sandwich ELISA, a second enzyme conjugated detecting antibody can then be added to bind the cytokine forming an antibody-antigen-antibody “sandwich”. The enzyme’s substrate is then added, producing spots (hence the name) of colour change within the culture. |
FLISA | Fluorescent Linked ImmunoSorbent Assays are a modification of ELISA assays that utilise fluorescent conjugated rather than enzyme conjugated antibodies to produce a detectable signal. |
ALISA | Aptamer Linked ImmunoSorbent Assays are a functional equivalent of ELISA that use aptamers rather than antibodies in either the capture or detection of antigen. |
ELONA | Enzyme Linked OligoNucleotide Assays are a functional equivalent of ELISA that use aptamers rather than antibodies in either the capture or detection of antigen. |
Gel Shift Assays | |
EMSA | An Electrophoretic Mobility Shift Assay separates protein-nucleotide complexes. |
Super Shift Assays | Super shift assays are an extension of EMSA, the addition of an antibody creates a greater shift but also allows identification of the protein present in the protein-DNA/RNA complex. |
Epitope Mapping | Epitope mapping is the process of identifying the target epitopes of an antibody within an antigen. |
Flow Cytometry | Cells in a liquid sample are passed one by one through a laser or intense light source. Data about the sample may be recovered from the direction and magnitude of light scatter. Fluorescent conjugated antibodies may also be used to label target proteins and allow identification and characterisation of cells. |
FACS | Fluorescent Activated Cell Sorting is an extension of flow cytometry that allows the subsequent sorting of cells. |
Immuno-electron Microscopy | Immuno-electron microscopy uses antibodies conjugated with heavy metals (commonly gold) to detect specific target proteins in tissues, which can then be visualised using transmission electron microscopy. |
Immunoaffinity-Mass Spectrometry | Immunoaffinity-mass spectrometry uses antibodies to enrich multiple proteins of interest from a sample, this is then followed by mass spectrometry-based quantification. |
Immunocytochemistry | Immunocytochemistry (ICC) is similar in concept to immunohistochemistry (IHC). Unlike IHC however, which allows visualisation of the distribution and localisation of specific components within a tissue: ICC is carried out on samples of intact cells lacking an extracellular matrix. |
Immunohistochemistry | Immunohistochemistry, often referred to as IHC, is a progression from histochemical analysis relying on highly specific antibody-antigen interactions as a method of protein detection. This allows visualisation of the distribution and localisation of specific cellular components within a cell or tissue. |
IHC-Frozen | Tissues are cryopreserved to limit ice crystal formation before embedding in OCT (optimal cutting temperature) cryostat medium. Sections of this embedded tissue are then cut using a cryomicrotome and immunohistochemical analysis performed. |
IHC-Paraffin | Tissues are embedded in paraffin blocks from which thin sections can be cut using microtome. immunohistochemical analysis can then be performed on these sections. |
IHC-Resin | Tissues are embedded in resin, commonly glycol methacrylate or methylmethacrylate allowing for thinner sections and less tissue shrinkage than might occur when using paraffin or cryo-embedding. |
IHC-Wholemount | Whole-mount immunohistochemistry involves probing intact tissues rather than sections. |
In Ovo | Experiments that are conducted in ovo are carried out within an egg. |
In Vitro | Experiments that are conducted in vitro are carried out “in glass” i.e. the entity to be examined is being viewed out with of it’s normal biological setting. |
In Vivo | Experiments that are conducted in vivo are carried out within a living organism. |
Inhibition Assays | |
Internalisation Experiments | |
Immunoprecipitation | Immunoprecipitation is a technique that uses immobilised antibodies specific to a target protein to precipitate out that protein from solution. The solid-phase substrate used to precipitate out antibody-antigen complexes can be attached to an antibody prior to being added to a sample. Alternatively beads coated in protein A/G capable of binding the antibody Fc region can be introduced once the antibody has already been added to a sample. |
ChIP | Chromatin Immunoprecipitation aims to determine whether individual proteins are associated with specific DNA sequences. Briefly, lengths of DNA are sheared down to smaller fragments and antibodies used to precipitate out those protein-DNA complexes of interest. The DNA can then be sequenced to determine the specific binding region of the associated binding protein. |
MeDIP | Methylated DNA immunoprecipitation utilises antibodies specific to 5-methylcytosine (5mC) to isolate and enrich for methylated DNA sequences. |
Luminex Assays | Luminex assays utilise antibody conjugated microspheres with unique fluorescent signatures to detect up to 500 different analytes within a sample. The fluorescent microsphere’s are detected using flow cytometric technology. |
MACS | Magnetic Activated Cell Sorting utilises antibodies with conjugated magnetic nanoparticles to separate cells. Cells are first incubated with a conjugated antibody directed at a particular surface antigen before being passed through a magnetic column. Those cells with an antibody conjugated to a magnetic nanoparticle remain in the column, while those without pass through. |
Neutralisation | |
Proximity Ligation Assays | Proximity ligation assays as the name suggests, rely on bringing into close proximity two DNA conjugated antibodies. While close to one another, these DNA sequences with the addition of further circle-forming DNA can be amplified via rolling circle amplification. Labeled complementary oligonucleotide probes can then be used to detect the amplified product. |
Protein-Lipid Overlay Assays | A Protein lipid overlay assay allows identification of the lipid ligands to which lipid binding proteins attach. |
Radioimmunoassays | Radioimmunoassays are a technique that can be used to measure the concentration of an antigen. Simply, a known quantity of antibody is mixed with a known quantity of its target antigen which has been radiolabeled. The sample containing an unknown concentration of antigen (unlabeled) is then added and the labeled and unlabeled antigen compete for antibody binding sites. The greater the concentration of unlabeled antigen present in the sample the more labeled antigen is unable to bind and remains free within the solution. This unbound labeled antigen can then be separated off and the radiation measured, a measurement from which we can then derive the concentration of unlabeled antigen present in the sample. |
Surface Plasmon Resonance | Surface plasmon resonance is a label-free technique capable of measuring antibody adsorption to an antigen coated plate, or antigen to an antibody coated plate. |
Biacore | Biacore assays utilise surface plasmon resonance based technology to measure antibody-antigen binding affinity. |
Thin Layer Chromatography – Enzyme Immunoassay | |
Western Blotting | A method for detecting the presence of specific proteins within a mixture. Samples are first separated by 3D structure (native proteins) or peptide length (denatured proteins) using electrophoresis before blotting to a membrane where they are then probed with antibodies specific to the target protein. |
Host
Host: The animal in which an antibody is raised (if polyclonal), or that provides the B cell component of a hybridoma (monoclonal).
Host | Description |
Chicken | Chickens are a commonly used host in the production of IgY, which can be collected and purified from their eggs. |
Mouse | Mice were the first species from which monoclonal antibodies were derived using a hybridoma. |
Goat | |
Rabbit |
Reactivity
Reactivity: The species from which an immunizing epitope was derived. Antibodies may also be reactive to other species depending on the sequence homology of the epitope.
*If you know of any resources that we have missed or that you would like to see included, please do drop Matt an email at matt@citeab.com
Reactive Species | Description |
Acanthamoeba | For more information on the genus Acanthamoeba, checkout AmoebaDB. |
Agricultural models | AgBase is a curated, open-source, Web-accessible resource for functional analysis of agricultural plant and animal gene products including Gene Ontology annotations. |
Arabidopsis thaliana | For more information on Arabidopsis thaliana, checkout The Arabidopsis Information Resource. |
Aspergillus | For more information on the genus Aspergillus, checkout The Aspergillus Genomes and the Aspergillus Cloud. |
Bacteria | Given their inextricable relationship with human and animal health and economic activity it’s no surprise that researchers and companies alike are producing antibodies to a vast array of bacterial and viral targets. CiteAb alone lists antibodies to over 160 different bacterial species of bacteriaincluding not only human pathogens but animal too. This translates to around 5,500 antibodies, with Salmonella and Streptococcus being the most popular genera. |
Caenorhabditis elegans | The nematode Caenorhabditis elegans is a model organism utilised in the study of neural development. More information on C. elegans and other worms and nematodes used in research can be found at WormBase. |
Candida | For more information on the genus Candida, checkout The Candida Genome Database. |
Cryptosporidium | For more information on the genus Cryptosporidium, checkout CryptoDB. |
Dictyostelium | Dictyostelium amoebae are model organisms commonly used in the study of cytokinesis, cell motility, phagocytosis, chemotaxis, signal transduction, and cell differentiation during development. They contain many genes that are homologous to those in higher eukaryotes yet are missing in Saccharomyces cerevisiae [dictyBase]. More information on Dictyostelium can be found at DictyBase. |
Drosophila | The small fruit fly Drosophila melanogaster, over the last century has become on of the most popular model organisms thanks in part to it’s short life cycle and the low cost and ease with which large numbers can be bred and housed. During this time it has been used extensively in the study of genetics and developmental biology. More information on Drosophila can be found at FlyBase. |
Entamoeba | For more information on the genus Entamoeba, checkout AmoebaDB. |
Escherichia coli | |
Epitope tag/Fusion protein | You can now add epitope tags and fusion proteins to your favourite protein, opening up a wide range of applications, from purifying your protein for structural studies to watching it function live within a cell. It is really useful to have an antibody for the tag or fusion protein you are using and some of the most cited antibodies in CiteAb are directed against common tags. Most companies from the largest to the smallest make antibodies to epitope tags or fusion proteins. |
Giardia lamblia | Giardia lamblia is a significant, environmentally transmitted, human pathogen. It is a valuable, if not unique, model for gaining basic insights into genetic innovations that led to formation of eukaryotic cells [GiardiaDB]. For more information on Giardia lamblia, checkout the Giardia Genomics Resource. |
Human | |
Mouse | Laboratory mice are the most widely used model organism on earth. |
Microsporidia | For more information on phylum Microsporidia, checkout MicrosporidiaDB. |
Piroplasma | For more information on the genus Piroplasma, checkout PiroplasmaDB. |
Plant | The large number of plant targeting antibodies is unsurprising given our age old dependency on plants as a source of food, fuel and materials for construction. Many companies produce, and some specialise in plant antibodies and CiteAb lists more than two and a half thousand antibodies capable of binding a range of plant species, all the way from Abies balsamea (Balsam fir) to Zea mays (Maize/Corn). |
Plasmodium | For more information on the genus Plasmodium, checkout PlasmoDB. |
Rat | |
Saccharomyces cerevisiae | The budding yeast Saccharomyces cerevisiae is a popular model system for the study of human disease genes. In 1996 it became the first eukaryote to have it’s entire genome sequenced. More information on S. cerevisiae can be found at the Saccharomyces Genome Database and their wiki; SGD-wiki. |
Schizosaccharomyces pombe | The fission yeast Schizosaccharomyces pombe is a model organism used in the study of the cell cycle and cell division. More info on S. pombe can be found at PomBase. |
Tetrahymena | For more information on Tetrahymena, checkout the Tetrahymena Genome Database and Wiki. |
Toxoplasma | For more information on genus Toxoplasma, checkout ToxoDB. |
Trypanosomatidae | For more information on the family Trypanosomatidae, checkout TritrypDB. |
Trichomonas | For more information on the genus Trichomonas and Trichomonas vaginalis in particular, checkout TrichDB. |
Virus | Given their inextricable relationship with human and animal health and economic activity it’s no surprise that researchers and companies alike are producing antibodies to a vast array of bacterial and viral targets. CiteAb alone lists almost 15,000 antibodies to 127 different viral species including not only human pathogens but animal too. |
Xenopus | The African clawed frog (Xenopus laevis), is a commonly used model organism in the study of embryonic development and early patterning events. More information on Xenopus laevis and X. tropicalis can be found at Xenbase. |
Zebrafish | Zebrafish (Danio rerio) are a commonly used model organism for vertebrate development and disease. More information about Zebrafish can be found at The Zebrafish International Resource Center (ZIRC) and The Zebrafish Information Network (ZFIN). |
Suppliers
There are many suppliers around the world that manufacture and distribute antibodies. This list, which is by no means exhaustive, provides a brief introduction to each supplier (In the interest of impartiality, each description is taken from the suppliers website and the list ordered alphabetically).
*This list is an ongoing effort and will be updated periodically.
Supplier | Description |
Abbexa | For more on Abbexa head over to our company highlight blog post, or alternatively you can access their website here. |
Abbiotec | |
Abbiotec | |
Abcam | |
ABclonal | |
Abcore | |
AbD Serotec | AbD Serotec antibodies may also appear in publications under the company names: Serotec and Bio-Rad |
Abgent | |
Abnova | |
Academic Antibodies (Individual labs) | CiteAb is able to list academic antibodies and we’re really keen to hear from those of you who have developed your own. To find out more about listing an antibody that you’ve raised in your lab, check out out blog post here. |
Academy Bio-medical | For more on Academy Bio-medical head over to our company highlight blog post, or alternatively you can access their website here. |
Acris Antibodies | |
Active Motif | |
Agrisera | |
Aldevron | Aldevron antibodies may also appear in publications under the company name Genovac. |
AntibodyBcn | |
Artron BioResearch | |
Atlas Antibodies | |
Aves Labs | |
Aviva Systems Biology | |
B-Bridge International | |
Badrilla | For more on Badrilla head over to our company highlight blog post, or alternatively you can access their website here. |
BD Biosciences | |
Beckman Coulter | |
Bethyl Laboratories | |
BioLegend | |
Biologo | For more on Biologo head over to our company highlight blog post, or alternatively you can access their website here. |
Biorbyt | |
Biosensis | |
Bioss | For more on Bioss head over to our company highlight blog post, or alternatively you can access their website here. |
Biotrend | |
Bioworld | |
Boster Immunoleader | |
Cell Signaling Technology | |
Covance | Covance antibodies may also appear in publications under the company names: Signet and Babco. |
Creative BioMart | |
Creative Diagnostics | |
Dako | |
DB Biotech | |
Developmental Studies Hybridoma Bank | Developmental Studies Hybridoma Bank antibodies may also appear in publications under the company’s abreviated name: DSHB. |
Diagenode | |
eBioscience | |
Echelon | |
EnCor Biotechnology | |
EnoGene | |
Enzo Life Sciences | Enzo Life Sciences antibodies may also appear in publications under the company name: Alexis Biochemicals. |
Epigentek | For more on Epigentek head over to our company highlight blog post, or alternatively you can access their website here. |
Epitope Biotech | |
Eurogentec | |
Everest Biotech | For more on Everest Biotech head over to our company highlight blog post, or alternatively you can access their website here. |
FabGennix | |
Fitzgerald | For more on Fitzgerald head over to our company highlight blog post, or alternatively you can access their website here. |
Gallus Immunotech | |
GeneTex | |
GenWay | |
Green Mountain Antibodies | |
Hycult Biotech | |
HyTest | |
IBT Bioservices | For more on IBT Bioservices head over to our company highlight blog post, or alternatively you can access their website here. |
ImmunoReagents | |
ImmunoStar | |
ImmuQuest | |
Indoor Biotechnologies | |
Jackson ImmunoResearch | For more on Jackson ImmunoResearch head over to our company highlight blog post, or alternatively you can access their website here. |
Kamiya Biomedical Company | |
Life Technologies | Life Technologies antibodies may also appear in publications under the company names: Invitrogen and Zymed. |
LSBio | |
Mab Technologies | |
Maine Biotechnology Services | |
MBL International | |
Médimabs | |
Millipore | Millipore antibodies may also appear in publications under the company names: Upstate, Chemicon and Merk Millipore. |
Miltenyi Biotec | |
Molecular Innovations | |
MUbio | |
NeuroMab | |
Nordic Immunological Laboratories | |
Nordic BioSite | |
Novus | Novus antibodies may also appear in publications under the company name Imgenex, which was acquired in 2013. |
PhosphoSolutions | For more on PhosphoSolutions head over to our company highlight blog post, or alternatively you can access their website here. |
Podiceps | |
Progen Biotechnik | For more on Progen Biotechnik head over to our company highlight blog post, or alternatively you can access their website here. |
ProSci | |
Proteintech | |
QED Bioscience | |
R&D Systems | |
Rockland Immunochemicals | |
Santa Cruz Biotechnology | |
Serascience | For more on Serascience head over to our company highlight blog post, or alternatively you can access their website here. |
SICGEN | |
Sigma-Aldrich | |
Signalway Antibody | For more on Signalway Antibody head over to our company highlight blog post, or alternatively you can access their website here. |
Sino Biological | |
Source BioScience | |
St John’s Laboratory | For more on St John’s Laboratory head over to our company highlight blog post, or alternatively you can access their website here. |
StressMarq Biosciences | For more on StressMarq Biosciences head over to our company highlight blog post, or alternatively you can access their website here. |
Synaptic Systems | |
TaKaRa | |
The European Malaria Reagent Repository | For more on The European Malaira Reagent Repository, head over to our company highlight blog post, or alternatively you can access their website here. |
Thermo Scientific | Thermo Scientific antibodies may also appear in publications under the company names: Lab Vision, NeoMarkers, ThermoFisher, ThermoPierce, Endogen and ABR Affinity Bioreagents. For more on Thermo Scientific head over to our company highlight blog post, or alternatively you can access their website here. |
Tonbo Biosciences | |
Trillium Diagnostics | |
US Biological | |
VIVA Bioscience | |
VMRD | VMRD antibodies may also appear in publications under the company’s full name: Veterinary Medical Research & Development. |
ZIRC | Zirc antibodies may also appear in publications under the company’s full name: Zebrafish International Resource Center. |
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