far western blotting


Summary: A method that is derived from western blotting (BLOTTING, WESTERN) and is used to detect protein-protein interactions. The blotted proteins are probed with a non-antibody protein which can then be tagged with a labeled antibody.

Top Publications

  1. Tamura M, Shiozaki I, Ono S, Miyano K, Kunihiro S, Sasaki T. p40phox as an alternative organizer to p47phox in Nox2 activation: a new mechanism involving an interaction with p22phox. FEBS Lett. 2007;581:4533-8 pubmed
    ..Thus, p40(phox) can substitute for p47(phox) in the activation, interacting with p22(phox) and p67(phox) through their specific regions. ..
  2. Kadam K, D Souza S, Bandivdekar A, Natraj U. Identification and characterization of oviductal glycoprotein-binding protein partner on gametes: epitopic similarity to non-muscle myosin IIA, MYH 9. Mol Hum Reprod. 2006;12:275-82 pubmed
    ..The results suggest that OGP binds to both gametes through its interaction with MYH9 through the non-glycosylated N-terminal conserved region of OGP, spanning the residues 11-137. ..
  3. Strom A, Diecke S, Hunsmann G, Stuke A. Identification of prion protein binding proteins by combined use of far-Western immunoblotting, two dimensional gel electrophoresis and mass spectrometry. Proteomics. 2006;6:26-34 pubmed
    ..Especially the application of this technique to investigate other neurodegenerative diseases is promising. ..
  4. Fautsch M, Vrabel A, Johnson D. The identification of myocilin-associated proteins in the human trabecular meshwork. Exp Eye Res. 2006;82:1046-52 pubmed
    ..Characterization of these potential interacting proteins may help to better understand the function of myocilin in the trabecular meshwork and aqueous outflow pathway. ..
  5. Naryzhny S, Lee H. Proliferating cell nuclear antigen in the cytoplasm interacts with components of glycolysis and cancer. FEBS Lett. 2010;584:4292-8 pubmed publisher
    ..Surprisingly, PCNA is also associated with six glycolytic enzymes that are involved in the regulation of steps 4-9 in the glycolysis pathway. ..
  6. Rudtanatip T, Withyachumnarnkul B, Wongprasert K. Sulfated galactans from Gracilaria fisheri bind to shrimp haemocyte membrane proteins and stimulate the expression of immune genes. Fish Shellfish Immunol. 2015;47:231-8 pubmed publisher
    ..In conclusion, this study demonstrated that the SG-stimulated immune activity in haemocytes is mediated, in part, through the LGBP, and IMD-NF-κB pathway. ..
  7. Li H, Huang X, Zeng Z, Peng X, Peng B. Identification of the interactome between fish plasma proteins and Edwardsiella tarda reveals tissue-specific strategies against bacterial infection. Int J Biochem Cell Biol. 2016;78:260-267 pubmed publisher
    ..Thus, our results provide a robust demonstration of both bacterial initiators and host receptors or interacting proteins to further explore infection and anti-infective mechanisms between hosts and microbes. ..
  8. Nakatomi A, Yazawa M. Identification and characterization of a novel calcineurin-binding protein in scallop testis. J Biochem. 2003;133:159-64 pubmed
    ..CaNBP75 was predominantly expressed in testis and ovary of scallop. Thus, CaNBP75 may modulate the physiological function of calcineurin in the testis and ovary of scallop, such as in spermiogenesis or meiosis. ..
  9. Zhou C, Zha X, Shi P, Zhao P, Wang H, Zheng R, et al. Nuclear hormone receptor BmFTZ-F1 is involved in regulating the fibroin heavy chain gene in the silkworm, Bombyx mori. Biochim Biophys Acta. 2016;1860:2529-2536 pubmed publisher
    ..These findings provide new insights into the mechanism of regulation of the silk protein gene. ..

More Information


  1. Coleman M, Miller K, Beernink P, Yoshikawa D, Albala J. Identification of chromatin-related protein interactions using protein microarrays. Proteomics. 2003;3:2101-7 pubmed
    ..This is the first demonstration of an interaction between RAD51B and histone proteins that may be important for the successful repair of DNA double-strand breaks. ..
  2. Kraiczy P. Identification and Characterization of Borrelia burgdorferi Complement-Binding Proteins. Methods Mol Biol. 2018;1690:95-103 pubmed publisher
    ..b>Far western blotting enables the detection of protein-protein interactions in vitro using cell lysates containing the prey ..
  3. Uno T, Nomura Y, Nakamura M, Nakao A, Tajima S, Kanamaru K, et al. Expression, purification and characterization of methyl DNA binding protein from Bombyx mori. J Insect Sci. 2005;5:8 pubmed
    ..This is the first report that insect MBD is phosphorylated and is present in adult tissues. These results suggest that bMBD2/3 plays important roles in the DNA methylation-specific transcription of Bombyx mori. ..
  4. Clinton S, Bina J, Hatch T, Whitt M, Miller M. Binding and activation of host plasminogen on the surface of Francisella tularensis. BMC Microbiol. 2010;10:76 pubmed publisher
    ..The ability of FT to acquire surface bound PLG that can be activated on its surface may be an important virulence mechanism that results in an increase in initial infectivity, survival, and/or dissemination of this bacterium in vivo. ..
  5. Locate S, Colyer J, Gawler D, Walker J. Annexin A6 at the cardiac myocyte sarcolemma--evidence for self-association and binding to actin. Cell Biol Int. 2008;32:1388-96 pubmed publisher
    ..Potential annexin A6 binding proteins in the sarcolemma fraction were identified using Far Western blotting. Two major annexin A6 binding proteins were identified as actin and annexin A6 itself...
  6. Lee S, Alani E. Analysis of interactions between mismatch repair initiation factors and the replication processivity factor PCNA. J Mol Biol. 2006;355:175-84 pubmed
    ..We favor a model in which PCNA acts as a scaffold for consecutive protein-protein interactions that allow for the coordination of MMR steps. ..
  7. Forsman M, Pääkkönen V, Tjäderhane L, Vuoristo J, Kallioinen L, Salo T, et al. The expression of myoglobin and ROR2 protein in Dupuytren's disease. J Surg Res. 2008;146:271-5 pubmed
    ..In this study, we showed for the first time the connection of ROR2 in Dupuytren's disease. ROR2 and myoglobin may play an important role in the pathophysiology of this disease. ..
  8. Sarkar S, Chelvarajan L, Go Y, Cook F, Artiushin S, Mondal S, et al. Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor. J Virol. 2016;90:3366-84 pubmed publisher
    ..These findings represent a significant advance in our understanding of the fundamental mechanisms associated with the entry of EAV into susceptible cells. ..
  9. Chow S, Yu C, Guy G. Sprouty2 interacts with protein kinase C delta and disrupts phosphorylation of protein kinase D1. J Biol Chem. 2009;284:19623-36 pubmed publisher
    ..By disrupting the capacity of PKCdelta to phosphorylate its cognate substrates, Spry2 may serve to modulate PKCdelta signaling downstream of receptor tyrosine kinases and to regulate the physiological outcome. ..
  10. George R, Chan H, Ahmed Z, Suen K, Stevens C, Levitt J, et al. A complex of Shc and Ran-GTPase localises to the cell nucleus. Cell Mol Life Sci. 2009;66:711-20 pubmed publisher
    ..The nuclear transport factor protein NTF2, which functions in cohort with Ran, was shown to form a complex with both RAN and Shc, suggesting a mechanism for Shc entry into the nucleus as part of a tertiary complex. ..
  11. Clyne M, Duggan G, Naughton J, Bourke B. Methods to Assess the Direct Interaction of C. jejuni with Mucins. Methods Mol Biol. 2017;1512:107-115 pubmed
    ..Here we describe a far Western blotting approach to show how C...
  12. Machida K, Mayer B, Nollau P. Profiling the global tyrosine phosphorylation state. Mol Cell Proteomics. 2003;2:215-33 pubmed
    ..We also describe ongoing efforts to modify and improve SH2 profiling, including the development of a multiplexed assay system that will allow high-throughput functional profiling of the tyrosine phosphoproteome. ..
  13. Chi R, Olenych S, Kim K, Keller T. Smooth muscle alpha-actinin interaction with smitin. Int J Biochem Cell Biol. 2005;37:1470-82 pubmed
  14. Roche J, Survery S, Kreida S, Nesverova V, Ampah Korsah H, Gourdon M, et al. Phosphorylation of human aquaporin 2 (AQP2) allosterically controls its interaction with the lysosomal trafficking protein LIP5. J Biol Chem. 2017;292:14636-14648 pubmed publisher
  15. Linz L, Liu S, Chougule N, Bonning B. In Vitro Evidence Supports Membrane Alanyl Aminopeptidase N as a Receptor for a Plant Virus in the Pea Aphid Vector. J Virol. 2015;89:11203-12 pubmed publisher
    ..This discovery is particularly important since the disruption of plant virus binding to such a receptor may enable the development of a nonchemical strategy for controlling aphid-vectored plant viruses to maximize food production. ..
  16. Pedrazzi G, Perrera C, Blaser H, Kuster P, Marra G, Davies S, et al. Direct association of Bloom's syndrome gene product with the human mismatch repair protein MLH1. Nucleic Acids Res. 2001;29:4378-86 pubmed
    ..We conclude that the hMLH1-BLM interaction is not essential for post-replicative MMR, but, more likely, is required for some aspect of genetic recombination. ..
  17. Edmondson D, Roth S. Identification of protein interactions by far Western analysis. Curr Protoc Cell Biol. 2001;Chapter 17:Unit 17.2 pubmed publisher
    b>Far Western blotting is a method for detecting protein-protein interactions. This technique utilizes a nonantibody protein probe to detect interacting proteins immobilized on a membrane support...
  18. Fukumoto S. [Immunologic tests: Echinococcosis]. Nihon Rinsho. 2005;63 Suppl 7:280-2 pubmed
  19. Yang H, Ivan M, Min J, Kim W, Kaelin W. Analysis of von Hippel-Lindau hereditary cancer syndrome: implications of oxygen sensing. Methods Enzymol. 2004;381:320-35 pubmed
  20. Cui S, Arosio D, Doherty K, Brosh R, Falaschi A, Vindigni A. Analysis of the unwinding activity of the dimeric RECQ1 helicase in the presence of human replication protein A. Nucleic Acids Res. 2004;32:2158-70 pubmed
    ..In addition, our far western, ELISA and co-immunoprecipitation experiments demonstrate that RECQ1 physically interacts with the 70 kDa subunit of hRPA and that this interaction is not mediated by DNA. ..
  21. Yeom J, Jeon C, Madsen E, Park W. In vitro and in vivo interactions of ferredoxin-NADP+ reductases in Pseudomonas putida. J Biochem. 2009;145:481-91 pubmed publisher
    ..This study demonstrates for the first time that Fprs in P. putida function as diaphorase, Fd/Fld reductases and determines their preferred redox partner in vivo and in vitro. ..
  22. Garg A, Barnes P, Porgador A, Roy S, Wu S, Nanda J, et al. Vimentin expressed on Mycobacterium tuberculosis-infected human monocytes is involved in binding to the NKp46 receptor. J Immunol. 2006;177:6192-8 pubmed
    ..ELISA and Far Western blotting showed that recombinant vimentin bound to a NKp46 fusion protein...
  23. Hara K, Shiota M, Kido H, Watanabe K, Nagata K, Toyoda T. Inhibition of the protease activity of influenza virus RNA polymerase PA subunit by viral matrix protein. Microbiol Immunol. 2003;47:521-6 pubmed
    ..These results suggest that M1 binds to and inhibits the amidolytic activity of PA. ..
  24. Klingelhofer J, Møller H, Sumer E, Berg C, Poulsen M, Kiryushko D, et al. Epidermal growth factor receptor ligands as new extracellular targets for the metastasis-promoting S100A4 protein. FEBS J. 2009;276:5936-48 pubmed publisher
    ..digital abstract: * MINT-7256556: EGF (uniprotkb:P01133) binds (MI:0407) to S100A4 (uniprotkb:P26447) by far western blotting (MI:0047) * MINT-7256512: BC (uniprotkb:P35070) binds (MI:0407) to S100A4 (uniprotkb:P26447) by far western ..
  25. Jadwin J, Curran T, Lafontaine A, White F, Mayer B. Src homology 2 domains enhance tyrosine phosphorylation in vivo by protecting binding sites in their target proteins from dephosphorylation. J Biol Chem. 2018;293:623-637 pubmed publisher
  26. Dong G, Kalifa R, Nath P, Babichev Y, Gelkop S, Isakov N. Crk adaptor proteins regulate CD3? chain phosphorylation and TCR/CD3 down-modulation in activated T cells. Cell Signal. 2017;36:117-126 pubmed publisher
    ..Furthermore, Crk-dependent increased phosphorylation of CD3? coincided with inhibition of TCR downmodulation, supporting a positive role for Crk adaptor proteins in TCR-mediated signal amplification. ..
  27. Parashar A, Colvin K, Bignell D, Leskiw B. BldG and SCO3548 interact antagonistically to control key developmental processes in Streptomyces coelicolor. J Bacteriol. 2009;191:2541-50 pubmed publisher
    ..coelicolor. The conservation of bldG and sco3548 in other streptomycetes demonstrates that this system is likely a key regulatory switch controlling developmental processes throughout the genus Streptomyces. ..
  28. Yoh S, Cho H, Pickle L, Evans R, Jones K. The Spt6 SH2 domain binds Ser2-P RNAPII to direct Iws1-dependent mRNA splicing and export. Genes Dev. 2007;21:160-74 pubmed
    ..Thus binding of Spt6 to Ser2-P RNAPII provides a cotranscriptional mechanism to recruit Iws1, REF1/Aly, and associated mRNA processing, surveillance, and export factors to responsive genes. ..
  29. Tanaka T, Abe Y, Inoue N, Kim W, Kumura H, Nagasawa H, et al. The detection of bovine lactoferrin binding protein on Trypanosoma brucei. J Vet Med Sci. 2004;66:619-25 pubmed
    ..to bind bovine LF (bLF) by using a fluorescence test, streptavidin-biotin (SAB) microplate analysis, and far Western blotting using a biotin-streptavidin system. We found that bLF bound to components of T...
  30. Terao Y, Okamoto S, Kataoka K, Hamada S, Kawabata S. Protective immunity against Streptococcus pyogenes challenge in mice after immunization with fibronectin-binding protein. J Infect Dis. 2005;192:2081-91 pubmed
    ..The Fn-binding RD and N terminus of FbaA are potential vaccine candidates for M1 strains of S. pyogenes infection. ..
  31. Zhou B, Liu J, Wang Q, Liu X, Li X, Li P, et al. The nucleocapsid protein of severe acute respiratory syndrome coronavirus inhibits cell cytokinesis and proliferation by interacting with translation elongation factor 1alpha. J Virol. 2008;82:6962-71 pubmed publisher
    ..Proliferation of human peripheral blood lymphocytes and other human cell lines was significantly inhibited by the infection of recombinant retrovirus expressing SARS-CoV N protein...
  32. Vepachedu V, Setlow P. Analysis of interactions between nutrient germinant receptors and SpoVA proteins of Bacillus subtilis spores. FEMS Microbiol Lett. 2007;274:42-7 pubmed
    ..Consequently, these new findings suggest that nutrient germinant receptors physically contact SpoVA proteins, and presumably this is a route for signal transduction during spore germination. ..
  33. Cootauco A, Murphy J, Maleski J, Blakemore K, SLODZINSKI M. Atrial natriuretic peptide production and natriuretic peptide receptors in the human uterus and their effect on myometrial relaxation. Am J Obstet Gynecol. 2008;199:429.e1-6 pubmed publisher
    ..The concentration of ANP may vary with gestational age and modulators of uterine contractility. NPR-A, -B, and -C receptor proteins are present in myometrial tissue. NPR-A levels may correlate with gestational age. ..
  34. Sheets A, Massey C, Cronk S, Iafrati M, Herman I. Matrix- and plasma-derived peptides promote tissue-specific injury responses and wound healing in diabetic swine. J Transl Med. 2016;14:197 pubmed publisher
    ..Comb1 labels keratinocytes, fibroblasts, and vascular endothelial cells in porcine wounds, and Far western blotting reveals these cell populations express multiple fluorescein-Comb1-interacting proteins in vitro...
  35. Feldman M, Zusman T, Hagag S, Segal G. Coevolution between nonhomologous but functionally similar proteins and their conserved partners in the Legionella pathogenesis system. Proc Natl Acad Sci U S A. 2005;102:12206-11 pubmed
    ..We hypothesize that this gene variation, which is probably mediated by positive selection, plays an important role in the evolutionary arms race between the protozoan host cell and the pathogen...
  36. Llorens F, Roher N, Miro F, Sarno S, Ruiz F, Meggio F, et al. Eukaryotic translation-initiation factor eIF2beta binds to protein kinase CK2: effects on CK2alpha activity. Biochem J. 2003;375:623-31 pubmed
    ..The results demonstrate that the ability to associate with CK2 subunits and to serve as a CK2 substrate are confined to different regions in eIF2beta and that it may act as an inhibitor on CK2alpha. ..
  37. Sun Y, Dai L, Sun Y, Wang L, Qian C, Wei G, et al. Gene expression patterns in response to pathogen challenge and interaction with hemolin suggest that the Yippee protein of Antheraea pernyi is involved in the innate immune response. J Invertebr Pathol. 2016;138:10-7 pubmed publisher
    ..Therefore, our data suggest that the Ap-Yippee protein is involved in a pathway that regulates the immune response of insects. ..
  38. Ghosh S, Thakur M. Interaction of estrogen receptor-alpha transactivation domain with nuclear proteins of mouse brain: p68 RNA helicase shows age- and sex-specific change. J Neurosci Res. 2009;87:1323-8 pubmed publisher
    ..Such age- and sex-specific interaction of p68 suggests its implication in ERalpha-mediated brain functions during aging...
  39. Fernandez Calvino L, Goytia E, López Abella D, Giner A, Urizarna M, Vilaplana L, et al. The helper-component protease transmission factor of tobacco etch potyvirus binds specifically to an aphid ribosomal protein homologous to the laminin receptor precursor. J Gen Virol. 2010;91:2862-73 pubmed publisher
    ..This variant retained its ability to bind CP but failed to interact with RPS2. ..
  40. Grote M, Wolf E, Will C, Lemm I, Agafonov D, Schomburg A, et al. Molecular architecture of the human Prp19/CDC5L complex. Mol Cell Biol. 2010;30:2105-19 pubmed publisher
    ..Our findings not only elucidate the molecular organization of the hPrp19/CDC5L complex but also provide insights into potential protein-protein interactions at the core of the catalytically active spliceosome. ..
  41. Liska F, Gosele C, Rivkin E, TRES L, Cardoso M, Domaing P, et al. Rat hd mutation reveals an essential role of centrobin in spermatid head shaping and assembly of the head-tail coupling apparatus. Biol Reprod. 2009;81:1196-205 pubmed publisher
    ..Cntrob may represent a novel candidate gene for presently unexplained hereditary forms of teratozoospermia and the "easily decapitated sperm syndrome" in humans. ..
  42. Wang W, Liu F, Chen N. [Effects of peroxisome proliferators-activated receptor gamma agonists on transforming growth factor-beta1 and Smads signal pathway: experiment with rat renal fibroblasts]. Zhonghua Yi Xue Za Zhi. 2006;86:740-4 pubmed
  43. Battesti A, Bouveret E. Improvement of bacterial two-hybrid vectors for detection of fusion proteins and transfer to pBAD-tandem affinity purification, calmodulin binding peptide, or 6-histidine tag vectors. Proteomics. 2008;8:4768-71 pubmed publisher
    ..Furthermore, we describe a set of vectors for TAP, CBP or 6-histidine tagging that possess the same cloning site as our two-hybrid vectors. ..
  44. Ren J, Lian T, Shao L, Liu Y, Liu Q. PmpI antibody reduce the inhibitory effect of Vp1 on Chlamydia trachomatis Infectivity. Can J Microbiol. 2018;: pubmed publisher
    ..trachomatis. Our research provides clues regarding the molecular mechanisms underlying the interactions between Chlamydia virus and Chlamydia. ..
  45. Zheng X, Yang M, Tan J, Pan Q, Long Z, Dai H, et al. Screening of LRRK2 interactants by yeast 2-hybrid analysis. Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2008;33:883-91 pubmed
    ..Our screened proteins may provide a new research clue for revealing biological functions of LRRK2, pathogenesis of Parkinson's disease, and other neurodegerations. ..
  46. Jahangiri A, Nguyen A, Chandra A, Sidorov M, Yagnik G, Rick J, et al. Cross-activating c-Met/?1 integrin complex drives metastasis and invasive resistance in cancer. Proc Natl Acad Sci U S A. 2017;114:E8685-E8694 pubmed publisher
  47. Tompkins K, George A, Veis A. Characterization of a mouse amelogenin [A-4]/M59 cell surface receptor. Bone. 2006;38:172-80 pubmed
  48. Zhang L, Zhao G, Hu X, Liu J, Li M, Batool K, et al. Cry11Aa Interacts with the ATP-Binding Protein from Culex quinquefasciatus To Improve the Toxicity. J Agric Food Chem. 2017;: pubmed publisher
    ..quinquefasciatus. Our study suggests that midgut proteins other than the toxin receptors may modulate the toxicity of Cry toxins against mosquitoes...
  49. Van de Casteele M, Omasta A, Janssens S, Roskams T, Desmet V, Nevens F, et al. In vivo gene transfer of endothelial nitric oxide synthase decreases portal pressure in anaesthetised carbon tetrachloride cirrhotic rats. Gut. 2002;51:440-5 pubmed
    ..These data indicate a major haemodynamic role of intrahepatic NOS 3 in the pathogenesis of portal hypertension in CCl(4) cirrhosis. ..
  50. Fecková B, Kimáková P, Ilkovičová L, Szentpéteriová E, Debeljak N, Solárová Z, et al. Far-western blotting as a solution to the non-specificity of the anti-erythropoietin receptor antibody. Oncol Lett. 2016;12:1575-1580 pubmed
    ..Furthermore, modified immunoprecipitation of EpoR followed by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry analysis confirmed a 57 kDa protein as a human Epo-interacting protein in both cell lines. ..
  51. Jadwin J, Mayer B, Machida K. Detection and quantification of protein-protein interactions by far-western blotting. Methods Mol Biol. 2015;1312:379-98 pubmed publisher
    ..We also present a batch quantification method that allows for the direct comparison of probe binding patterns. ..
  52. Kristufek D, Aspalter R, Eibl M, Wolf H. Characterization of novel Bruton's tyrosine kinase gene mutations in Central European patients with agammaglobulinemia. Mol Immunol. 2007;44:1639-43 pubmed
    ..The results of this study further substantiate the importance of using elaborate molecular analysis with different detection techniques to obtain an explicit molecular diagnosis in patients with suspected XLA. ..
  53. Sun R, Zhao X, Wang Z, Yang J, Zhao L, Zhan B, et al. Trichinella spiralis Paramyosin Binds Human Complement C1q and Inhibits Classical Complement Activation. PLoS Negl Trop Dis. 2015;9:e0004310 pubmed publisher
    ..The binding of recombinant and natural Ts-Pmy to human C1q were determined by ELISA, Far Western blotting and immunoprecipitation, respectively...