Qian P, Wang X, Zhong CQ, Wang J, Cai M, Nguitragool W, Li J, Cui H, Yuan J. Elife. as a chromogenic substrate, according to the following protocol. Objective: This study aimed to determine the diagnostic efficiency of a novel immunoblotting detection assay for anti-ganglioside antibodies (AGAs) in the Guillain-Barre syndrome (GBS).Method: Serum immunoglobulin (IgG and IgM) of AGAs were measured in 121 participants from a registered cohort study of immune-mediated neuropathies and 29 healthy controls by immunoblotting panel assay . Tissue-specific expression pattern and iden-tification of functional cis-and trans-regulatory elements. Anal. A new membrane substrate for western blotting of proteins. Epub 2013 Mar 5. After washing off any unbound primary antibody, the membrane is incubated with a secondary antibody that recognizes and binds to the primary antibody. HHS Vulnerability Disclosure, Help Similarly, the protein content of the samples can be compared ("sample A contains more protein than sample B"). detection technique where an immunoblot is initially tested with polyclonal antibodies . The western blot (sometimes called the protein immunoblot ), or western blotting, is a widely used analytical technique in molecular biology and immunogenetics to detect specific proteins in a sample of tissue homogenate or extract. Yamamoto T, Matsuo T, Yamamoto A, Yamagoshi R, Ohkura K, Kataoka M, Shinohara Y. Methods 276, 98389845. , clinical, and forensic medicine applications. Therefore the following separation happens solely by the size of the polypeptide chains in the polyacrylamide gel. When the ligand is not an antibody, the reaction can be visualized using a ligand that is directly labeled. The size separation, prior to blotting, allows the protein molecular weight to be gauged, as compared with known molecular weight markers. (1984) Calmodulin-binding proteins: visualization by 125I-Calmodulin overlay on blots quenched with Tween 20 or bovine serum albumin and poly(ethylene oxide). In Western blotting (immunoblotting) the protein mixture is applied to a gel electrophoresis in a carrier matrix (SDS-PAGE, native PAGE, isoelectric focusing, 2D gel electrophoresis, etc.) 2022 Apr 12;4:100109. doi: 10.1016/j.fochms.2022.100109. A disadvantage of western blotting (immunoblot) is that it is time-consuming (compared to ELISA) and has a high demand in terms of experience of the experimenter. Yes, there are alternative protein analysis methods such as ELISA, mass spectrometry, protein arrays, flow cytometry, and immunohistochemical techniques that can be applied depending on the application. (1982) The use of Tween 20 as a blocking agent in the immunological detection of pro-teins transferred to nitrocellulose membrane. Anal. Methods Biochem. 21, 841845. 145, 315321. In clinical diagnostic settings, immunoelectrophoresis is applied, which involves the electrophoresis of serum or urine followed by immunodiffusion. The method is based on building an antibody:protein complex via specific binding of antibodies to proteins immobilized on a membrane and detecting the bound antibody with one of several detection methods. Glenney, J. The stacking gel has a low percentage of acrylamide and serves to concentrate and focus the protein sample into a narrow band, while the resolving gel has a higher percentage of acrylamide and separates the proteins based on their size.For reducing Western blotting, a reducing agent such as beta-mercaptoethanol or dithiothreitol (DTT) is often added to the sample buffer to break the disulfide bonds between cysteine residues and fully denature the proteins. Bernstein, D. I., Garraty, E., Lovett, M. A., and Bryson, Y. J. Pasquali, C., Vilbois, F., Curchod, M.-L., van Huijsduijnen, R. H., and Arigoni, F. (2000) Mapping and identification of protein-protein interactions by two-dimensional far-western immunoblotting. 33, 158. CAS Proteomics. They involve identification of protein target via antigen-antibody (or protein-ligand) specific reactions. The immunoblotting technique or western blotting is used to detect proteins of interest in a complex mixture of proteins in different cellular or tissue samples using specific . Anal. Clipboard, Search History, and several other advanced features are temporarily unavailable. - 167.99.76.245. Protein binding to the membrane is an irreversible mechanism. . Eur. Membranes can be of the nitrocellulose, polyvinylidene difluoride (PVDF), or nylon variety. Root, D. D. and Reisler, E. (1989) Copper iodide staining of protein blots on nitrocellulose membranes. Western blotting is a versatile technique that allows detection of a particular protein in a sample. Biochim. Western Blotting (also called immunoblotting) is a technique used for analysis of individual proteins in a protein mixture (e.g. FEMS Immunol. Gooley, A. A 7-Step Guide to Western Blotting | Cytiva The technique involves a series of steps that begin with the separation of proteins based on their size using gel electrophoresis. Unwin, R. D., Harnden, P., Pappin, D., Rahman, D., Whelan, P., Cra-ven, R. A., et al. Western blotting (immunoblot) expands on the idea of ELISA by allowing separation of the protein mix by size, charge, and/or conformation. Biochem. Electrophoresis For best experience we recommend to activate Javascript in your browser. Membranes can be of the nitrocellulose, polyvinylidene difluoride (PVDF), or nylon variety. CrossRef The PubMed wordmark and PubMed logo are registered trademarks of the U.S. Department of Health and Human Services (HHS). The separated protein bands are then transferred to a carrier membrane (e.g. Acad. J. Immunol. Please enable it to take advantage of the complete set of features! Sadra, A., Cinek, T., and Imboden, J. Dot blot is a simplified procedure in which protein samples are not separated by electrophoresis but are spotted directly onto membrane. Physiol. 55, 297307. anticorps-enligne.fr, chinese () Hsp70-Bag3 Module Regulates Macrophage Motility and Tumor Infiltration via Transcription Factor LITAF and CSF1. Hawkes, R., Niday, E., and Gordon, J. This technique exploits the specificity inherent in antigen-antibody recognition. Electrophore-sis Careers. Anal. Chem. J. Biochem. It is also one of the standard techniques for characterization of antibodies from different samples of polyclonal sera or hybridoma supernatants. J Epilepsy Res. The site is secure. Janke, C., Holzer, M., Klose, J, and Arendt, T. (1996) Distribution of isoforms of the microtubule-associated protein tau in grey and white matter areas of human brain: a two-dimensional gelelectrophoretic analysis. Anal. Copyright 2023 by Cold Spring Harbor Laboratory Press. Biochem. Bookshelf 21, 33573368. Anal. Unable to load your collection due to an error, Unable to load your delegates due to an error. Additionally, SDS binds the proteins and thereby covers their chemical charges, leading to equally negatively charged proteins. Copyright 2023 by Cold Spring Harbor Laboratory Press. Immunoblotting can be performed using the antibodies raised against synthetic peptide antigens modified Int. In immunoblot techniques such as Western blot analysis, proteins are separated by electrophoresis and transferred onto nitrocellulose sheets, then are identified by their reaction with labeled antibodies. They involve identification of protein target via antigen-antibody (or protein-ligand) specific reactions. Immunoblotting has a variety of research, clinical, and forensic medicine applications. A method of analysis in tissue extracts. 123, 229235. 13, 197203. Immunoblotting Techniques | Request PDF - ResearchGate Methods They mixture is then rapidly cooled down on ice before loading to keep proteins in their denatured, linear form. 262, 10,03510,038. Depending on the pH the acidic and basic functional groups contribute by increasing or decreasing the total charge of the protein. The .gov means its official. Proteins are generally separated by size using sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis. This method allows for the separation of proteins that are inaccessible by other methods. Biochem. A primary antibody specific for the protein of interest or alternatively a protein tag is used for binding to the protein. Electrophoresis In a tank blot (wet transfer), the gel is sandwiched between two pads or sponges soaked in transfer buffer, and a membrane such as nitrocellulose or PVDF is placed on top of the gel. This site needs JavaScript to work properly. Michalewski, M. P., Kaczmarski, W., Golabek, A., Kida, E., Kaczmarski, A., Wisniewski, K. E. (1999) Immunoblotting with antiphosphoamino acid antibodies: importance of the blocking solu-tion. Nielsen, P. J., Manchester, K. L., Towbin, H., Gordon, J., and Tho-mas, G. (1982) The phosphorylation of ribosomal protein S6 in rat tissues following cycloheximide injection, in diabetes, and after den-ervation of diaphragm. It is commonly used in molecular biology to determine protein expression and detect the target proteins in biological samples. Wilkinson, D. (2000) Chemiluminescent techniques for western blot detection let researcher shed their lead aprons. Proteomics Hong, H. Y., Yoo, G. S., and Choi, J. K. (2000) Direct Blue 71 staining of proteins bound to blotting membrane. The antibodies are conjugated with fluorescent or radioactive labels or enzymes that give a subsequent reaction with an applied reagent, leading to a coloring or emission of light, enabling detection. Sci. A simple SDS-PAGE immunoblotting technique using an enhanced 257, 1231612321. Ikegaki, N. and Kennett R. H. (1989) Glutaraldehyde fixation of the primary antibody-antigen complex on nitrocellulose paper increases the overall sensitivity of immunoblot assay. Anal. From the Abstract Book of 2nd Siena 2-D Electrophoresis Meeting, 57; Siena Sept. 16-18. 131, 115. Natural passive immunity. J. Immunol. Immunoblotting of 2-DE Separated Proteins | SpringerLink Dunn, S. D. (1986) Effects of the modification of transfer buffer com-position and the renaturation of proteins in gels on the recognition of proteins on western blots by monoclonal antibodies. CrossRef Immunoblotting techniques use antibodies (or other specific ligands in related techniques) to identify target proteins among a number of unrelated protein species. It is based on the principle of immunochromatography where proteins are separated into polyacrylamide gel according to their molecular weight. Western blotting allows for high specific protein identification, semi-quantitative analysis, the ability to examine multiple proteins in one sample, and depending on the experimental layout the detection of protein interactions. This method can also detect different complexes of different proteins. (1997) An expression cloning method to identify monomeric GTP-binding proteins by GTP over-lay. 16, 11901192. The membrane can then be blocked with serum albumin or milk solution to prevent non-specific antibody binding. Reinhart, M. P. and Malamud, D. (1982) Protein transfer from iso-electric focusing gels: the native blot. A. Murray, J., Gilkerson, R., and Capaldi, R. A. 270, 270283. Gershoni, J. M. and Palade, G. E. (1982) Electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to a posi-tively charged membrane filter. (2000) Multiple probing of an immunoblot membrane using a non-block technique: advantages in speed and sensitivity. in order to detect other specific antibodies from the protein mixture under investigation). Some of the main applications of Western blotting are disease diagnosis by detecting specific markers, antibody validation for downstream applications such as immunohistochemistry or flow cytometry, detecting antibodies in serum samples to assess vaccine efficacy, or quality control in biosimilars production to ensure consistency. 251, 6972. It accounts for 80% of rhinitis in children and 30% in adults. Immunoblots are most often used in research settings and are usually performed to confirm results from ELISA or other immunoassays. ), Springer-Verlag Berlin Heidelberg. Szewczyk, B. and Kozloff, L. M. (1985) A method for the efficient blotting of strongly basic proteins from sodium dodecyl sulphate-polyacrylamide gels to nitrocellulose. Methods Schloss-Rahe-Str. Kamps, M. P. (1991) Generation of anti-phosphotyrosine antibodies for immunoblotting. 156, 341347. The immunoblotting (western blotting) has been useful in identifying specific antigens recognized by polyclonal or monoclonal antibodies and is highly sensitive. 15, 223230. Biotin-conjugated antibodies are also used for this purpose. Chem. Their structure and possibly interactions with other proteins remain intact and influence the migration in during electrophoresis. 2015;1312:487-503. doi: 10.1007/978-1-4939-2694-7_51. 14, 2932. Western Blotting (Immunoblot): Gel Electrophoresis for Proteins Biochem. Ch. 17 - Immunization and Diagnostic Immunoassays Flashcards 26, 85398545. Gallagher S, Winston SE, Fuller SA, Hurrell JG. In situ PLA, applied on traditional immunoblotting membranes, has been shown to improve detection specificity through the use of pairs of antibodies, as well as detection sensitivity through the . Reproducible results across biological and technical replicates ensure the quality of the data. For a reducing Western blot, samples are dissolved in a Laemmli buffer and then boiled to assure that all the protein in the sample is denatured. Big proteins migrate slower and are represented by the highest bands on the blot, while small proteins migrate faster and are indicated by the lowest bands on the blot. Methods J. Biol. It is typically performed under non-reducing conditions, meaning that the disulfide bonds between cysteine residues are not reduced. eCollection 2022 Jul 30. Native-native 2D gel electrophoresis for HDL subpopulation analysis. Through a series of chemical reactions, the bound antibodies are visualized, allowing for the identification and quantification of the protein. An official website of the United States government. to mimic posttranslational modifications of proteins, such as phosphorylation and acetylation, to study these modifications To this end, the electrophoretically separated proteins in the sample are compared with a molecular weight standard of a known composition. Unable to load your collection due to an error, Unable to load your delegates due to an error. This analytic technique proceeds in the following steps. J. Immunol. Stott, D. I. National Library of Medicine COVID-19 Diagnostic Methods and Detection Techniques - PMC The technique identifies one or multiple protein targets at a time through two major steps, which are SDS-PAGE and immunoblotting, and it involves three key elements: (a) separation by size, (b) transfer to an adsorbent membrane and (c) identification of the target protein using the appropriate primary antibody, followed by secondary antibody tr. Biophys. eCollection 2016 Dec. Han S, Jerome JA, Gregory AD, Mallampalli RK. Methods Enzymol. Following the separation of the protein mix the polypeptide bands are transferred to a membrane carrier. FEBSLett. 2013 May;162(1):9-23. doi: 10.1104/pp.113.214460. Lemkin, P. (1996) Matching 2-D gels on the Internet. The described method of stripping allows for the detection of several targets, contrary to ELISA where only one protein can be detected. The site is secure. A western blot, sometimes called a protein immunoblot, is an antibody-based technique used to detect the presence, size and abundance of specific proteins within a sample. Immunoblotting Techniques | Springer Nature Experiments PubMed Preparing Immunoprecipitations for Immunoblotting. Immunoblotting can be performed using the antibodies raised against synthetic peptide antigens modified to mimic posttranslational modifications of proteins, such as phosphorylation and acetylation, to study these modifications in the protein of interest in vivo. Burnette, W. N. (1981) Western blotting: electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to un-modified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal. This is a preview of subscription content, access via your institution. Additionally, unspecific binding pockets can be blocked before the addition of specific antibodies. The proteins are then transferred onto a membrane and subsequently probed with specific antibodies that bind to the target protein of interest. Anal. Anal. This manual summarizes the most commonly used techniques, provides information about the wide selection of blotting apparatus and detection reagents . Western blotting (immunoblotting): history, theory, uses, protocol and problems | BioTechniques Login to your account Email: Password Anti-RNP antibodies are associated with mixed connective tissue disease, which may be considered a distinct clinical entity associated with a specific . Immunoblot assays are usually performed to confirm results obtained by other techniques such as ELISA. Hence, it covers very different topics and applications. 4, 272282. 2016 Dec 31;6(2):45-50. doi: 10.14581/jer.16010. 2. a cell lysate). Langstein, J. and Schwarz, H. (1997) Suppression of irrelevant sig-nals in immunoblots by preconjugation of primary antibodies. 19, 20102013. Reducing Western blots are commonly used to detect a particular protein or subunit of a protein complex. Pathol. Would you like email updates of new search results? Methods In Molecular Biology, vol 295. J. Biochem. Denaturing SDS-PAGE is typically used for separating proteins based on their molecular weight. 1, 890898. However, the applied electric charge causes the proteins to travel out of the gel vertically to the direction they traveled in on the gel, onto the membrane. 173, 1121. Curr. 291, 12571260. Stott, D. I. The immunoblot technique is a rapid and sensitive assay for the detection and characterization of proteins that works by exploiting the specificity inherent in antigen-antibody . The term 'protein expression' is often applied to describe immunohistochemical results, but it should be remembered that the technique only reflects localization of a protein, not the related RNA synthesis, and the gene coding for it may not be active . Anti-Sm antibodies are highly specific for SIE, being found in 76% of SLE patients. 2020 Cold Spring Harbor Laboratory Press, Alert me when Updates/Comments are published, Artistic rendition of adult male and female killifish. This analytic technique proceeds in the following steps. 251, 288291. Methods Mol Biol. (1994) Mapping of Ras-related GTP-binding pro-teins by GTP overlay following two-dimensional gel electrophoresis. Therefore, it can be applied to a wide range of research and clinical applications, making it an essential tool for many scientists and clinicians. Electrophoresis Biochemical approach for isolation of polyadenylated RNAs with bound proteins from yeast. A. Gao, Y., Wells, L., Comer, F. I., Parker, G. J., and Hart, G. W. (2001) Dynamic O-glycosylation of nuclear and cytosolic proteins: cloning and characterization of a neutral, cytosolic beta-N-acetylglucosaminidase from human brain. Daneels, G., Moeremans, M., De Raeymaeker, M., and De Mey, J. Immunoblotting allows detection of a protein antigen immobilized on the protein-retaining membrane support such as nitrocellulose or polyvinylidene fluoride (PVDF). clinical, and forensic medicine applications. Proteins are typically separated by electrophoresis and transferred onto membranes (usually nitrocellulose). cellular fractions. Bookshelf Immunoblotting technique, also dubbed "Western blotting," exploits antibody specificity to identify target proteins among a number of unrelated protein species (1,2).Traditionally used for mapping known proteins on electrophoretically-resolved mixtures, in the proteome era immunoblotting has been combined with 2-D gel electrophoresis for rapid visualization and identification of target . Poxton, I. R. (1990) Immunoblotting techniques. 19, 752757. Immunoblotting is now widely used in conjunction with two-dimensional polyacrylamide gel electrophoresis, not only for traditional goals, such as the immunoaffinity identification of proteins and analysis of immune responses but also as a genome-proteome interface technique. B. Immunoblotting can be used as a highly sensitive method for detection of anti-Sm and anti-RNP antibodies. Typical problems in Western blotting include non-specific binding and background signals, weak signal and difficulty in detecting lowly expressed proteins, and variability between experiments. Natl. Google Scholar. or polyvinylidene fluoride (PVDF). The "blotted" bands are now available to be treated further (e.g. 8600 Rockville Pike Immunoblotting allows detection of a protein antigen immobilized on the protein-retaining membrane support such as nitrocellulose Due to its upper and lower detection limits Western blot is primarily a qualitative method. 2017;1554:61-73. doi: 10.1007/978-1-4939-6759-9_3. Accessibility StatementFor more information contact us atinfo@libretexts.org. A labelled protein which binds to DNA or RNA of interest in order to visualize it Methods Methods Lin, W. and Kasamatsu, H. (1983) On the electrotransfer of polypep-tides from gels to nitrocellulose membrane. Western Blot Procedures, Analysis and Purpose Altmetric, Part of the Methods In Molecular Biology book series (MIMB,volume 295). Immunology Chapter 20 Quiz Flashcards | Quizlet Sanchez-Campillo, M., Bini, L., Comanducci, M., Raggiaschi, R., Marzocchi, B., Pallini, V., et al. Immunohistochemistry has an essential role in the evaluation of muscle biopsies and in examining protein localization. Immunoblotting - ResearchGate and transmitted securely. Big proteins migrate slower and are represented by the highest bands on the blot, while small proteins migrate faster and are indicated by the lowest bands on the blot. Electrophoresis (1985) Comparison of western blot analysis to microneutralization for the detection of type-specific antibodies to herpes simplex virus antibod-ies. { "12.2A:_Immunoassays_for_Disease" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "12.2B:_Antibody_Functions" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "12.2C:_Serology" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "12.2D:_Precipitation_Reactions" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "12.2E:__Agglutination_Reactions" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "12.2F:_Neutralization_Reaction" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "12.2G:_Complement_Fixation" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "12.2H:_Fluorescent_Antibodies" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "12.2I:_Enzyme-Linked_Immunosorbent_Assay_(ELISA)" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "12.2J:_Immunoblot_Procedures" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "12.2K:_Tests_That_Differentiate_Between_T_Cells_and_B_cells" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "12.2L:__In_Vivo_Testing" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "12.2M:_The_Future_of_Diagnostic_Immunology" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()" }, { "12.01:_Immunization" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "12.02:_Immunoassays_for_Disease" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "12.03:_Preparations_for_Diagnosing_Infection" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "12.04:_Immunity_Disorders-_Hypersensitivity" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "12.05:_Immunity_Disorders-_Autoimmune_Diseases" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "12.06:_Immunity_Disorders-_Immunodeficiencies" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()" }, [ "article:topic", "showtoc:no", "license:ccbysa", "licenseversion:40" ], https://bio.libretexts.org/@app/auth/3/login?returnto=https%3A%2F%2Fbio.libretexts.org%2FBookshelves%2FMicrobiology%2FMicrobiology_(Boundless)%2F12%253A_Immunology_Applications%2F12.02%253A_Immunoassays_for_Disease%2F12.2J%253A_Immunoblot_Procedures, \( \newcommand{\vecs}[1]{\overset { \scriptstyle \rightharpoonup} {\mathbf{#1}}}\) \( \newcommand{\vecd}[1]{\overset{-\!-\!\rightharpoonup}{\vphantom{a}\smash{#1}}} \)\(\newcommand{\id}{\mathrm{id}}\) \( \newcommand{\Span}{\mathrm{span}}\) \( \newcommand{\kernel}{\mathrm{null}\,}\) \( \newcommand{\range}{\mathrm{range}\,}\) \( \newcommand{\RealPart}{\mathrm{Re}}\) \( \newcommand{\ImaginaryPart}{\mathrm{Im}}\) \( \newcommand{\Argument}{\mathrm{Arg}}\) \( \newcommand{\norm}[1]{\| #1 \|}\) \( \newcommand{\inner}[2]{\langle #1, #2 \rangle}\) \( \newcommand{\Span}{\mathrm{span}}\) \(\newcommand{\id}{\mathrm{id}}\) \( \newcommand{\Span}{\mathrm{span}}\) \( \newcommand{\kernel}{\mathrm{null}\,}\) \( \newcommand{\range}{\mathrm{range}\,}\) \( \newcommand{\RealPart}{\mathrm{Re}}\) \( \newcommand{\ImaginaryPart}{\mathrm{Im}}\) \( \newcommand{\Argument}{\mathrm{Arg}}\) \( \newcommand{\norm}[1]{\| #1 \|}\) \( \newcommand{\inner}[2]{\langle #1, #2 \rangle}\) \( \newcommand{\Span}{\mathrm{span}}\)\(\newcommand{\AA}{\unicode[.8,0]{x212B}}\), 12.2I: Enzyme-Linked Immunosorbent Assay (ELISA), 12.2K: Tests That Differentiate Between T Cells and B cells, Describe how Western blotting allows individuals to detect specific solubilized proteins from serum or cell or tissue extracts.