Whole mount antibody staining was performed according to the zebrafish book with slight modifications 63. Applications key ab array antibody array ae affinity electrophoresis affin chroma. Prior to labeling with the antibody, the embryos or larvae must be fixed. Investigating microglia during motor neuron degeneration using a zebrafish model. The mechanistic target of rapamycin mtor is a serinethreonine protein kinase that regulates cell growth, cell proliferation, protein synthesis, and autophagy. The mammalian target of rapamycin mtor partner, raptor. Ubtorkiaa1024 regulates neurite outgrowth and neoplasia. Intriguingly, rapamycin, an inhibitor of mtor, can delay cyst. It crossreacts weakly with some other proteins based on western analysis. As the leading manufacturer of zebrafish antibodies, genetex offers an extensive portfolio of reagents validated for use in this model system. This antibody revealed mtor to be at subcellular structures very similar to those i had failed to define earlier.
Mammalian target of rapamycin mtor is a conserved serthr kinase that is part of mtor complex 1 mtorc1, a master regulator that couples amino acid availability to cell growth and autophagy. The mtor antibody from proteintech is a rabbit polyclonal antibody to a peptide of human mtor. Golph3 protein localizes to the golgi network and its overexpression stimulates s6 ribosomal kinase 1, which is activated by mtor, part of the pi3kakt mtor. Using both the zebrafish intersegmental vessel and hyaloid vessel assays to. About these antibodies, view the anatomical ontology browser complete list of. Role of insulin receptor signaling in zebrafish embryogenesis. Feel free to add new protocols to the appropriate section or add comments to any existing protocol. Zebrafish is an ideal model for the study of neurogenesis. Ulkatgfip200 complexes mediate mtor signaling to the. The pi3kaktmtor pathway is a complex signalling pathway. Protocols are organized into sections corresponding to the chapters of the zebrafish book, 5th edition 4th edition online. Anti mtor phospho s2448 antibody epr4262 bsa and azide free ab177734 description. Golph3 modulates mtor signalling and rapamycin sensitivity. Zebrafish danio rerio, a minnowlike freshwater fish, has emerged as a popular animal model for research and development.
To investigate the role of mtorc1 signaling pathway in zebrafish caudal fin, we examined the activation and function of mtorc1 signaling using an antibody against phosphorylated s6. The mtor antibody from cell signaling technology is a rabbit polyclonal antibody to frap, frap2, and mtor. Zebrafish international resource center zirc china zebrafish resource center czrc. Our tormtor antibodies can be used in a variety of model species. Our understanding of the mechanisms involved in cancer genesis and progression underwent unprecedented expansion during the last decade, opening a new era of. Our understanding of the mechanisms involved in cancer genesis and progression underwent unprecedented expansion during the last decade, opening a new era of cancer treatment targeted therapy. Human orthologs of this gene implicated in autosomal dominant polycystic kidney disease.
These include methods and techniques used for the study of the mtor pathway and potential therapeutic applications of mtor inhibitors such as, immunosuppressive and anticancer agents. Intraspinal sensory neurons provide powerful inhibition to motor circuits. Expression of ztorassociated micrornas in zebrafish. The mtor antibody has been validated for the following applications. The mammalian target of rapamycin mtor controls multiple cellular functions in response to amino acids and growth factors, in part by regulating the phosphorylation of p70 s6 kinase p70s6k and eukaryotic initiation factor 4ebinding protein 1 4ebp1. Zpr1 antibody can combine with protein encoded by arrestin3a arr3a. Nod1 deficiency impairs cd44alck as well as pi3kakt pathway.
Rabbit monoclonal epr4262 to mtor phospho s2448 bsa and azide free. Zebrafish as a model to understand autophagy and its role. Moreover, since our knowledge in this area rapidly evolves, some data have got obsolete during the process of book editing. The mechanism of mtor mammalian target of rapamycin in a mouse model of polycystic ovary syndrome pcos. Methods and protocols methods in molecular biology. B the 3d larvae labeled with antiacetylated tubulin antibody brown, lateral view with anterior to the left. Mechanistic target of rapamycin complex 1 signaling regulates cell. Active mechanistic target of rapamycin plays an ancillary rather than. Genetex currently offers a continuously expanding catalog of reagents designed expressly for zebrafish research, with. We appreciate as much detail as possible and references as appropriate. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding ser2481 of human mtor.
It is also unclear if ztor is involved in similar biological functions in the zebrafish even though it is an orthologue of mtor. Today, zfin provides the official nomenclature for zebrafish genes, mutants. We employed two pcr arrays to profile the pi3kakt and mtor. Due to their striking genomic similarity to humans, regenerative abilities, and shortened life cycle, zebrafish have become an ideal tool for studying developmental processes, gene expression, and underlying mechanisms of cancer and other diseases. Deciphering combinations of pi3kaktmtor pathway drugs. Zebrafish antibodies primary antibodies sigmaaldrich.
Tormtor antibody used for ihcp on tissue specimens obtained from patients having carcinomas of the gallbladder gbca, extrahepatic bile duct carcinoma ehbdca and intrahepatic bile duct carcinoma ihbdca. Use the list below to choose the tor mtor antibody which is most appropriate for your research. Sigma life sciences offers over 300 zebrafish antibodies representing research areas such. This is where zebrafish researchers can share information with the rest of the community. In the present study, we demonstrated that suppression of ztor is associated with physiological changes, including reduced embryonic senescence and melanogenesis and decreased angiogenic response. These results clearly demonstrate that nod1 antibody specifically detect endogenous nod1 protein in zebrafish right in fig. If you continue, well assume that you are happy to receive all cookies. As for mtor kinase assay, mtor or myctagged immunoprecipitates were obtained from 293t cells using anti mtor antibody santa cruz biotechnology, sc1549 or antimyc antibody 9e10 or a purified, active form of mtor millipore, bedford, ma was used. Dot blot analysis of mtor phospho s2448 phospho peptide lane 1 and mtor nonphospho peptide lane 2 labeling mtor phospho s2448 phospho peptide with purified ab109268 at a dilution of 1 0. Immunoblotting with a monoclonal anti mtor antibody detected mtor protein at comparable levels in both es cells and hek cells fig.
Immunohistochemical analysis of paraffinembedded zebrafish tissue, using mtor gtx124771 antibody at 1. Zirc monoclonal antibodies zebrafish international. Mitochondrial control of apoptosis 47 mtor signaling 45 nfkb signaling 29. Immunohistochemistry of wholemount and sectioned zebrafish tissue. Genes related to inflammation and immune response were upregulated in tsc2. Article pdf available in journal of ovarian research 51. Hela cells were transfected with either cy3 dye labeled negative control sirna nc or mtor sirna mtor sikd and then stained with the antibody against mtor. As a part of the pi3kakt pathway, there is a broad interest in mtor biology across cancer types. This is where zebrafish researchers can share experimental protocols and tips with the rest of the research community.
A immunofluorescence signal was reduced by sirna mediated knockdown of mtor protein. The monoclonal antibodies were generated with balbc mice against adult zebrafish protein, and clones were selected based on the specific staining patterns obtained with particular clones. A 28h wholemount zebrafish embryo labeled with an antibody which recognizes the transcription factor, pax6 brown label showing a lateral view with anterior to the left. Cell lysates were collected from human embryonic kidney hek cells and es cells. Zebrafish were acclimated for 15 days and fed twice daily with a commercial diet. Mtor antibody detects mtor protein on zebrafish by whole mount immunohistochemical analysis. As substrates, atg, ulk1 651end, and ulk2 651end were purified from e. We also monitored mtor expression at the protein level by western blot analysis. The protein encoded by this gene belongs to a family of phosphatidylinositol kinaserelated kinases. Zebrafish larvae lacking depdc5 show mtor hyperactivation, seizures. Antibody information for antibodies hpa071227 and cab069425 used in analysis of ensg00000198793 mtor flj44809, frap, frap1, frap2, raft1, rapt1 we use cookies to enhance the usability of our website. The mtor antibody confirms silencing of mtor expression, and the eif4b antibody is used to control for loading and sirna specificity.
Monoclonal antibody 92 polyclonal antibody 21 modification. More excitingly, he and others in the laboratory found that within minutes of starving cells for amino acids or glucose, mtor left the punctate structures and became diffuse throughout the cytoplasm 53, 55. Please use this form to recommend updates to the information in zfin. Transcriptomics revealed 1414 differentially expressed genes between wt and tsc2. Zebrafish international resource center zirc order this. Treatment of zebrafish with rapamycin during early embryogenesis resulted in developmental delay, but did not cause any overt defects. The main objective of this book is to provide an uptodate survey of the rapidly advancing eld of cancer therapy.
Methods and protocols expert researchers in the field detail many of the methods which are now commonly used to study mtor. Orthologous to human mtor mechanistic target of rapamycin kinase. Zebrafish have a single homologue of mtor which, although expressed ubiquitously during early embryogenesis, becomes localized to the head and developing gut between 35 and 57 hours postfertilization h. Two tsc1 homologs were identified in zebrafish, which we refer to as tsc1a and tsc1b. Ihc assay involved antigen retrieval with 10 mm sodium citrate buffer ph 6 employing high power of microwave treatment for 10 minutes and. Conversely, overexpression of cd44a in nod1deficient zebrafish.
Antibody 2972 mtor antibody by cell signaling technology. Full event breakdown with abstracts, speakers, registration and more. The mtor substrates antibody sampler kit provides an economical means to evaluate the signaling of mtor to downstream substrates including p70 s6 kinase and 4ebp1. Zebrafish tsc1 reveals functional interactions between the cilium. Noncanonical mtorindependent role of depdc5 in regulating. The mammalian target of rapamycin mtor, sometimes also referred to as the mechanistic target of rapamycin and fk506binding protein 12rapamycinassociated protein 1 frap1, is a kinase that in humans is encoded by the mtor gene. Our antibody search, biochemicals search and kits and assays search provides a simple way for researchers to find reagents that work, while our high quality market data and citations are used commercially by many of the worlds leading reagent suppliers and investment companies. The hybridomas are stored at the monoclonal antibody facility of the uo under the supervision of dr. Knockout of zebrafish interleukin 7 receptor il7r by the crispr.
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