Though more expensive, the PVDF membrane is stronger and has a high absorption capacity with proteins, which makes it suitable for CBB staining, detection with certain substrates for alkaline phosphatase, and re-probing. Nitrocellulose and PVDF membranes are commonly used. Comparison between blotting apparatuses Membrane A semi-dry or a tank system can be used for transfer. Proteins separated by SDS-PAGE are “transferred” from the polyacrylamide gel to a membrane, using a specialized apparatus (blotting apparatus). The principle and method of polyacrylamide gel electrophoresis (SDS-PAGE).The following sections describe the procedure starting from electrotransfer of proteins to the membrane.įollow the links below for the methods used to generate antibodies, and the principle and method of SDS-PAGE. The bound enzyme activity is used to detect the target protein and visualized by a chemiluminescent or chromogenic method. The membrane with transferred proteins is then probed with a primary antibody (an antibody specific for the target protein), washed, and reacted with a secondary antibody labeled with an enzyme, such as horseradish peroxidase (HRP). In Western blotting (WB), target proteins are transferred to a hydrophobic membrane after SDS-PAGE and detected using specific antibodies.Īfter SDS-PAGE, a membrane is placed on the gel, to which the separated proteins in the gel are electrophoretically transferred. FAQs – In Vivo Isotype Control AntibodiesĪdvanced MHC Tetramers and Antibodies Searchįind products for Western blot The principle.Protein-Protein Interaction Detection Technology.FGF-Max (Recombinant human FGF-1/FGF-2 chimera).Afamin/Wnt3a CM and recombinant protein Show submenu.Non-Classical MHC Tetramers Show submenu.MHC Class I & Class II Tetramers Show submenu.SARS-CoV-2 MHC-Peptide Screening Kit Bundles.QuickSwitch™ Custom Tetramer Kits+ Show submenu.Custom MHC Tetramers and Monomers Order Form.Krohn RI (2011) The colorimetric detection and quantitation of total protein.

Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Walhout AJ (2006) Biochemistry and molecular biology. Nat Protoc 9:375–395įonslow BR, Moresco JJ, Tu PG, Aalto AP, Pasquinelli AE, Dillin AG, Yates JR 3rd (2014) Mass spectrometry-based shotgun proteomic analysis of C. PLoS One 5:e10161Īrur S, Schedl T (2014) Generation and purification of highly specific antibodies for detecting post-translationally modified proteins in vivo. Hadwiger G, Dour S, Arur S, Fox P, Nonet ML (2010) A monoclonal antibody toolkit for C. Kim HM, Colaiacovo MP (2016) CRISPR-Cas9-guided genome engineering in C. Nat Methods 11:529–534ĭickinson DJ, Goldstein B (2016) CRISPR-based methods for Caenorhabditis elegans genome engineering. Nat Genet 40:1375–1383įrokjaer-Jensen C, Davis MW, Sarov M, Taylor J, Flibotte S, LaBella M, Pozniakovsky A, Moerman DG, Jorgensen EM (2014) Random and targeted transgene insertion in Caenorhabditis elegans using a modified Mos1 transposon. Methods Cell Biol 106:161–185įrokjaer-Jensen C, Davis MW, Hopkins CE, Newman BJ, Thummel JM, Olesen SP, Grunnet M, Jorgensen EM (2008) Single-copy insertion of transgenes in Caenorhabditis elegans. Praitis V, Maduro MF (2011) Transgenesis in C. Methods Cell Biol 107:67–92Įvans TC (2005) Transformation and microinjection.

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Western blot analysis