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Niswender CM, Conn PJ (2010) Metabotropic glutamate receptors: physiology, pharmacology, and disease.Mameli M et al (2007) Rapid synthesis and synaptic insertion of GluR2 for mGluR-LTD in the ventral tegmental area.Balschun D et al (1999) A specific role for group I mGluRs in hippocampal LTP and hippocampus-dependent spatial learning.Bortolotto ZA et al (1994) A molecular switch activated by metabotropic glutamate receptors regulates induction of long-term potentiation.Conn PJ, Pin JP (1997) Pharmacology and functions of metabotropic glutamate receptors.Bass JJ et al (2017) An overview of technical considerations for Western blotting applications to physiological research.Murphy RM, Lamb GD (2013) Important considerations for protein analyses using antibody based techniques: down-sizing Western blotting up-sizes outcomes.Coorssen JR et al (2002) Quantitative femto- to attomole immunodetection of regulated secretory vesicle proteins critical to exocytosis.Huang Q et al (2014) A non-radioactive method for small RNA detection by northern blotting.Fields C et al (2019) Northern blot with IR fluorescent probes: strategies for probe preparation.Curr Protoc Mol Biol Chapter 12:Unitas 12.8 Papoulas O (2001) Rapid separation of protein-bound DNA from free DNA using nitrocellulose filters.Pall GS, Hamilton AJ (2008) Improved northern blot method for enhanced detection of small RNA.Damm K et al (2015) Improved northern blot detection of small RNAs using EDC crosslinking and DNA/LNA probes.Anderson GJ, Cipolla CM, Kennedy RT (2011) Western blotting using capillary electrophoresis.Wilhelm BG et al (2014) Composition of isolated synaptic boutons reveals the amounts of vesicle trafficking proteins.Gallo RC et al (1984) Frequent detection and isolation of cytopathic retroviruses (HTLV-III) from patients with AIDS and at risk for AIDS.Burnette 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.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.Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.Ninfa AJ, Ballou DP, Benor M (2009) Fundamental laboratory approaches for biochemistry and biotechnology, 2nd edn.Gibbons J (2014) Western blot: protein transfer overview.Science/AAAS Custom Publishing Office, Athens Perkel JM (2015) A history of the old West(ern blot).While other immunoblotting methods can also be used, this chapter contains general information that will aid researchers in the ability to conduct western blot experiments, especially those relating to mGlu receptor expression. The overarching goal of this chapter is to discuss the theory of western blotting, applying of this technique for answering scientific questions regarding changes in mGlu receptor protein expression, and provide a standardized western blotting protocol for immunoblotting of these receptors. In addition, mGlu receptors frequently form heterodimers or homodimers with other receptors. However, the promiscuity of mGlu receptor antibodies has yielded inconsistent results with high levels of nonspecific binding and background staining. Identifying proteins of interest, such as metabotropic glutamatergic receptors (mGlu receptors), has proven beneficial for addressing key questions within the field.
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This technique has been used for over four decades and remains a popular method for both protein detection and quantification. Western blotting is a highly adaptable biochemical assay that is used widely across cellular and molecular biology disciplines and has a diverse array of applications.