Western blot analysis of extracts from various mouse tissues using 3. Pillai-Kastoori L, Schutz-Geschwender AR, Harford JA (2020) A systematic approach to quantitative Western blot analysis. The results are summarized in the reference table. Sci Signal 8:rs2Ĭlarke DC, Liu X (2010) Measuring the absolute abundance of the Smad transcription factors using quantitative immunoblotting. Janes KA (2015) An analysis of critical factors for quantitative immunoblotting. McDonough AA, Veiras LC, Minas JN, Ralph DL (2015) Considerations when quantitating protein abundance by immunoblot. Hill CS (2016) Transcriptional control by the SMADs. Shi Y, Massague J (2003) Mechanisms of TGF-beta signaling from cell membrane to the nucleus. A Defined Methodology for Reliable Quantification of Western Blot Data. Gene editing in human stem cells using zinc. Massague J (1998) TGF-beta signal transduction. The western blot image was exported to ImageJ software to measure band density. Van der Kraan PM (2017) The changing role of TGFbeta in healthy, ageing and osteoarthritic joints. Nature 425:737–741Ĭolak S, Ten Dijke P (2017) Targeting TGF-beta Signaling in cancer. Ghaemmaghami S, Huh WK, Bower K, Howson RW, Belle A, Dephoure N, O'Shea EK, Weissman JS (2003) Global analysis of protein expression in yeast. 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. Crit Rev Biochem Mol Biol 51:110–119Ĭhong YT, Koh JL, Friesen H, Duffy SK, Cox MJ, Moses A, Moffat J, Boone C, Andrews BJ (2015) Yeast proteome dynamics from single cell imaging and automated analysis. Four Simple Steps for Western Blot Quantification 1. So on to the four critical steps for ensuring your (semi) quantification is accurate. Torres NP, Ho B, Brown GW (2016) High-throughput fluorescence microscopic analysis of protein abundance and localization in budding yeast. Even though western blot quantification is only semi-quantitative, you still need to be rigorous in how you quantify your blots to ensure your results are as accurate as possible. Annu Rev Anal Chem (Palo Alto, Calif) 11:49–77 Annu Rev Anal Chem (Palo Alto, Calif) 9:449–472Īnkney JA, Muneer A, Chen X (2018) Relative and absolute quantitation in mass spectrometry-based proteomics. Gillet LC, Leitner A, Aebersold R (2016) Mass spectrometry applied to bottom-up proteomics: entering the high-throughput era for hypothesis testing. Nat Cell Biol 8:1195–1203īeck M, Schmidt A, Malmstroem J, Claassen M, Ori A, Szymborska A, Herzog F, Rinner O, Ellenberg J, Aebersold R (2011) The quantitative proteome of a human cell line. Nature 499:79–82Īldridge BB, Burke JM, Lauffenburger DA, Sorger PK (2006) Physicochemical modelling of cell signalling pathways. Wu L, Candille SI, Choi Y, Xie D, Jiang L, Li-Pook-Than J, Tang H, Snyder M (2013) Variation and genetic control of protein abundance in humans. Gnann C, Cesnik AJ, Lundberg E (2021) Illuminating non-genetic cellular heterogeneity with imaging-based spatial proteomics. BMC Biology 14, Article number: 116 ( 2016 ) Cite this article. Ho B, Baryshnikova A, Brown GW (2018) Unification of protein abundance datasets yields a quantitative Saccharomyces cerevisiae proteome. Quantifying western blots: none more black.
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