The arlb sco coding sequence was amplified via pcr from a zebrafish. The encoded protein is a small gtpase that contains both nterminal and c. Using this antibody, we detected the presence of arlb on cilia in all of the. Because tropical aquarium fish are not used for food purposes, they pose no threat to the food supply. Gfp expression while cilia structural mutations resulting in shortened cilia are detectable by antibody staining as described above, the ability to specifically detect cilia paralysis or normal immotile cilia in vivo can be difficult as this requires live imaging of nonmoving axonemes. Analysis of cilia structure and function in zebrafish.
The arl8b antibody 1a2 from novus biologicals is a mouse monoclonal antibody to arl8b. A guide for the laboratory use of zebrafish danio rerio. To further investigate the role of arlb in ciliogenesis, we examined cilia in cultured wildtype and arlb hnn mefs. Monoclonal antibodies that recognize a variety of structures in zebrafish embryos are available from the library at the zebrafish international resource center. The cellular localization is predicted to be cytoplasmic. Bertram, in fetal and neonatal physiology fifth edition, 2017. Shield sample from light with aluminum foil from this step on. Zebrafish danio rerio, a minnowlike freshwater fish, has emerged as a popular animal model for research and development. Tested in western blot wb and immunohistochemistry paraffin ihc p applications. Defects in arlb are the cause of joubert syndrome js which is an autosomal recessive disorder characterized by a distinctive cerebellar malformation pmid. Select zebrafish protein antibodies from monoclonal antibodies listed below. Hrp conjugate antibody cell signaling at a dilution of 1.
This antibody also detected two nonspecific bands of lower molecular weight fig. Apr 28, 2020 from ncbi gene this gene encodes a member of the adpribosylation factorlike family. Abcam antibodies and reagents supplier, find any antibody. Nggcep164, a novel centriole appendage protein required for. Membrane enriched extracts 30 g lysate of hela lane 1, jurkat lane 2, k562 lane 3, hek293 lane 4, panc1 lane 5, mcf7 lane 6, hep g2 lane 7, mouse. Gfptagged murine arlb under the strong ubiquitous bactin2 promoter. 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. Thus, the development and characterization of mutant andor knock out zebrafish are critical to help define immune cell and immune gene functions in the zebrafish model. The arlbmcherry transgene was designed with a murine adpribosylation factorlike b arlb cdna fused inframe upstream of a monomeric red fluorescent protein, mcherry, and driven by the pcaggs promoter which contains a chicken. Online help keyboard shortcuts feed builder whats new. Arlb regulates ciliogenesis and the dynamic localization of shh signaling proteins christine e. In december 2005, a cancer research team headed by keith cheng at penn state university announced that their studies of the mutant golden zebrafish had.
Characterization of rag 1 mutant zebrafish leukocytes. The arl8b antibody 1a2 has been validated for the following applications. The more basic the biology is, the more choice you have in which model to study it in. Arlb silencing leads to reduced cd1a surface expression and function. In reciprocal experiments, protein extracts were immunoprecipitated. Involved in cilium assembly and photoreceptor cell outer segment organization.
The transgene was microinjected into fertilized fvb oocytes and founder mice were bred to. Increased support for mutants and transgenics article pdf available in nucleic acids research 41d1 october 2012 with 9 reads how we measure. Zebrafish antibodies primary antibodies sigmaaldrich. Is the zebrafish research community using a subset of. The hybridomas are stored at the monoclonal antibody facility of the uo under the supervision of dr. 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. Arlb has 3,347 functional associations with biological entities spanning 8 categories molecular profile, organism, disease, phenotype or trait, functional term, phrase or reference, chemical, structural feature, cell line, cell type or tissue, gene.
Fusion protein amino acids 208427 cterminus of mouse arlb also known as adpribosylation factorlike protein b, adpribosylation factorlike protein 2like 1, arl2like protein 1 and arl2l1, accession number. Fusion protein amino acids 208427 cterminus of mouse arlb also known as adpribosylation factorlike protein b, adpribosylation factorlike protein 2like 1, arl2like protein 1 and arl2l1, accession number q640n2 rat. View detailed zebrafish protein antibody specifications by linking to the specific product blocks. Hardcopies of the 4th edition of the zebrafish book can be obtained for a nominal fee from the zebrafish international resource center, 5274 university of oregon, eugene, or 97403 usa. This target gene encodes the protein adp ribosylation factor like gtpase b in humans and may also be known as arl2l1, jbts8, adpribosylation factorlike protein b, and adpribosylation factorlike b. Moreover, cilia were present on endothelial cells in the developing zebrafish vasculature. Arlb regulates ciliogenesis and the dynamic localization of. Arlb is predicted to be a 48 kda protein, and the 60 kda band is likely to represent a modified form of arlb. This antibody reacts with human,mouse,rat, dog samples. Anti arlb antibody products anti arlb antibodies are offered by a number of suppliers. There is no evidence that these genetically engineered zebra danio fish pose any more threat to the environment than their unmodified counterparts which have long been widely sold in the united states. Arlb antibody rabbit polyclonal from proteintech validated in western blot wb, immunoprecipitation ip, immunohistochemistry ihc, immunofluorescence if, enzymelinked immunosorbent assay elisa applications. Anderson home office, animals scientific procedures inspectorate, uk e.
These cells are commonly used for studies focusing on the biochemical and molecular properties of zebrafish. The commonly used antiacetylated tubulin monoclonal antibody 611b1 sigma t6793 works well to visualize cilia in zebrafish. C, zebrafish heterozygous for scohi459, a null allele of the zebrafish. Guidance on the housing and care ofzebrafishdanio rerio. For the most current and complete antibody data, use the antibody search page to see zirc antibodies only, set the search to show only zirc antibodies. Endothelial cilia are essential for developmental vascular integrity.
We previously demonstrated that arlb is expressed in cilia and does not overlap with the basal body in fibroblasts caspary et al. The encoded protein is a small gtpase that contains both nterminal and cterminal guanine nucleotidebinding motifs. Invitae is a college of american pathologists capaccredited and clinical laboratory improvement amendments cliacertified clinical diagnostic laboratory performing fullgene sequencing and deletionduplication analysis. Orthologous to human arlb adp ribosylation factor like gtpase b. Monoclonal, polyclonal, and recombinant antibodies. Moreover, the antibody recognizes the truncated cc2d2a protein. The library included five hairpins per gene and covered 211 genes. Arlb regulates ciliogenesis and the dynamic localization. The original zebrafish or zebra danio, danio rerio is a native of rivers in india and bangladesh. Mutations in the arlb gene are associated with the joubert syndrome. Find and compare multiple sources of anti antibody using the linscotts directory search engine. Zhiyuan gong and his colleagues at the national university of singapore were working with a gene that encodes the green fluorescent protein gfp, originally extracted from a jellyfish, that naturally produced. Zebrafish models for ciliopathies scoarlb, ift88 and ift57 mutants 811.
Select appropriate zebrafish protein antibodies for your research by isotype, epitope, applications and species reactivity. Small aliquots of antisco can be requested from the authors and a detailed protocol for immunostaining and imaging has been described elsewhere drummond, 2009. Guidance on the housing and care of zebrafish, danio rerio 3 acknowledgements the authors would like to thank the following people for their helpful comments during the preparation of this resource. Western blot analysis of arlb using 50 g of mouse liver lysate. In december 2005, a cancer research team headed by keith cheng at. Ftm is a novel basal body protein of cilia involved in shh signalling. The information on this page was automatically extracted from online scientific databases. As the vast majority of commercial antibodies are raised against human proteins, we became interested in taking a look at which commercial antibodies are being used by the zebrafish research community. Is the zebrafish research community using a subset of specialist antibodies raised for zebrafis. Primary antibodies used were arlb protein tech, 177111ap. Custom antibody development and commercial partnerships to advance your diagnostic and therapeutic discovery. Select applications, conjugates, hosts, and reactivity. Arlb function was demonstrated previously in zebrafish 15. The pronephros is the first of the excretory organs to form as the nephric duct differentiates.
Antibodies, primary antibodies, zfish zebrafish, alphabetical listing of zebrafish antibodies. The specificity of the arlb antibody was confirmed by immunoprecipitating extracts with the arlb antibody and immunoblotting with the same antibody fig. Effects of lethal irradiation in zebrafish and rescue by. Please feel free to communicate with us via email if you have any questions. Each dish was wrapped in parafilm then placed atop a 3inch spacer to achieve a dose rate of 978 cgymin in a 7 cesium source irradiator. Please use this form to recommend updates to the information in zfin. Characterization of rag 1 mutant zebrafish leukocytes bmc. We appreciate as much detail as possible and references as appropriate. The zebrafish, a small but flashy aquarium pet, may seem like an unlikely informant on questions of human genetics yet its genome could hold the keys to understanding many diseases and, surprisingly, the genes underlying human skin color. In addition, we have developed a rabbit polyclonal antiscorpion arlb antisco antibody that is also a robust and specific marker for cilia duldulao et al. 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. In reciprocal experiments, protein extracts were immunoprecipitated with antibodies against arlb and immunoblotted with antivangl2 antibodies. We assembled an shrna library targeting members of protein families known to be involved in membrane trafficking, including rabs, arfsarls, snares, escrt, tetraspanins, and adaptor protein complexes, among others. Anti fish antibody products listed below are anti fish antibodies from multiple suppliers.
Arlb rabbit anticanine, human, mouse, rat, polyclonal. Tested in western blot wb and immunohistochemistry ihc applications. The arlb gene encodes an axonemal protein that controls cilia length, and mutations in this gene lead to dysfunctional cilia pmid. Overexpression of arlb increases cilia length in zebrafish embryos in a. This protein is localized in the cilia and plays a role in cilia formation and in maintenance of cilia clinical significance. In humans, as with other amniotes, this is a transitory structure, which appears as 5 to 7 paired pronephroi that connect to the nephric duct around 3 weeks gestation. Palmitoylation of the ciliary gtpase arlb is necessary for its. Lysis of the frozen adult fish with 700 l np40 buffer including protease inhibitor using a tissue lyser ii and a metal bead for 2 times, 2 min, 25 hz. A function for the joubert syndrome protein arlb in ciliary. Arlb has 3,347 functional associations with biological entities spanning 8 categories molecular profile, organism, disease, phenotype or trait, functional term, phrase or reference, chemical, structural feature, cell line, cell type or tissue, gene, protein or microrna extracted from 68 datasets.
Antitlr9 in, zfishtm 55351 custom antibody service antibody labeling custom assay development zfishtmembryonic lysate 24 hpf 55347 zfishtm embryonic. If youre studying cytoskeletal dynamics, you can literally go. This antibody detects two specific bands at 60 kda and 48 kda. Acute irradiation injury was replicated using animal models, and seminal experiments by jacobsen et al 2,3 and lorenz et al 4 showed that shielding or. Zebrafish protein antibody scbt santa cruz biotechnology. Listed below are antifish antibodies from multiple suppliers. Fish is a reported alias name for the human gene sh3pxd2a, or sh3 and px domains 2a. It measures three centimeters long and has gold and dark blue stripes.
Antiarlb antibody products antiarlb antibodies are offered by a number of suppliers. Cilia localization is essential for in vivo functions of the joubert. Research and development of antibody production using. Kahnc, and tamara casparya adepartment of human genetics, school of medicine, bgraduate. Finally, the membrane was observed by developing with diaminobenzidine dab. Purified antiarginase 1 antibody antiarginase 1 o94e6arg1. About these antibodies all antibodies listed on this page were generated by researchers at the institute of neuroscience, university of oregon liu et al. The 13amino acid protein is a member of the sh3pxd2 family. Arlb antibody 6f11 is a mouse monoclonal igg 2a kappa light chain provided at 100 gml.
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