1/2/2024 0 Comments Western blot results of fish![]() ![]() We recommend using a wide-field fluorescence microscope though other imaging platforms can be used with Stellaris RNA FISH probes. The sample is ready for imaging using a standard fluorescence microscope. The total time for this step is 1-1.5 hours. Removal of Hybridization Buffer and addition of wash buffers with short incubation periods are able to remove excess probes. Hybridization can be completed in 4 to 16 hours at +37 ☌ in a standard laboratory incubator. Note: Variations in sample preparation between different organisms and sample types are covered in our online Stellaris protocols. For some sample types, the entire process can be completed in less than a day.Ī sample is grown, adhered, or sectioned onto a #1 cover glass and permeabilised with alcohol. Proc Natl Acad Sci U S A 76(9):4350–4354īradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.The Stellaris RNA FISH protocol is simple, consisting of four main steps, as shown in the image below. Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. ![]() Renart J, Reiser J, Stark GR (1979) Transfer of proteins from gels to diazobenzyloxymethyl-paper and detection with antisera: a method for studying antibody specificity and antigen structure. Laemmli UK (1970) Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4. J Proteome Res 9(9):4393–4406īurnette WN (1981) “Western Blotting”: electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Front Immunol 5:1–8Ĭarrera M, Cañas B, Vázquez J et al (2010) Extensive de Novo sequencing of new parvalbumin isoforms using a novel combination of bottom-up proteomics, accurate molecular mass measurement by FTICR-MS, and selected MS/MS ion monitoring. Kuehn A, Swoboda I, Arumugam K et al (2014) Fish allergens at a glance: variable allergenicity of parvalbumins, the major fish allergens. Pazos M, Méndez L, Vázquez M et al (2015) Proteomics analysis in frozen horse mackerel previously high-pressure processed. Pazos M, Méndez L, Gallardo JM et al (2014) Selective-targeted effect of high-pressure processing on proteins related to quality: a proteomics evidence in atlantic mackerel ( Scomber scombrus). Ortea I, Rodríguez A, Tabilo-Munizaga G et al (2010) Effect of hydrostatic high-pressure treatment on proteins, lipids and nucleotides in chilled farmed salmon ( Oncorhynchus kisutch) muscle. J Agric Food Chem 66(17):4559–4570Įtienne M, Jérôme M, Fleurence J et al (2000) Identification of fish species after cooking by SDS-PAGE and urea IEF: a collaborative study. J Food Eng 262:83–91Ĭarrera M, Fidalgo LG, Saraiva JA et al (2018) Effects of high-pressure treatment on the muscle proteome of hake by bottom-up proteomics. Tsironi T, Anjos L, Pinto PIS et al (2019) High pressure processing of European sea bass ( Dicentrarchus labrax) fillets and tools for flesh quality and shelf life monitoring. Mol Cell Proteomics 11(6):1–17Īnjos L, Pinto PIS, Tsironi T et al (2019) Experimental data from flesh quality assessment and shelf life monitoring of high pressure processed European sea bass ( Dicentrarchus labrax) fillets. Gillet LC, Navarro P, Tate S et al (2012) Targeted data extraction of the MS/MS spectra generated by data-independent acquisition: a new concept for consistent and accurate proteome analysis. FAOĪnjo SI, Santa C, Manadas B (2015) Short GeLC-SWATH: a fast and reliable quantitative approach for proteomic screenings. J Proteome 147:119–124įAO - Food and Agriculture Organization of the United Nations (2014) The state of world fisheries and aquaculture: opportunities and challenges. Mazzeo MF, Siciliano RA (2016) Proteomics for the authentication of fish species. Food Res Int 54(1):972–979Ĭarrera M, Pazos M, Gasset M (2020) Proteomics-based methodologies for the detection and quantification of seafood allergens. Carrera M, Cañas B, Gallardo JM (2013) Proteomics for the assessment of quality and safety of fishery products. ![]()
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