The zebrafish information network provides just as thorough of a resource on zebrafish husbandry for free. Zebrafish, danio rerio, were maintained and embryos were collected essentially. The zebrafish international resource center is a nonprofit organization. Phenotypic chemical screening using a zebrafish neural. Orthologous to several human genes including krt8 keratin 8. Panel d is immunoprecipitation of mycmmp17b and rfpreck in cos7 cells. It is aimed to help labome visitors find the most suited rfpred fluorescent proteindsredmcherry. Collections of plasmids that encode zebrafish receptors or monoclonal antibodies to zebrafish cell surface. The hybridomas are stored at the monoclonal antibody facility of the uo under the supervision of dr. B the 3d larvae labeled with antiacetylated tubulin antibody. Biotin conjugated polyclonal anti rfp used in a sandwich elisa with unconjugated anti rfp is well suited titrate rfp.
Zebrafish danio rerio is a fresh water fish that inhabits rivers in india. Zebrafish antibodies primary antibodies sigmaaldrich. Zebrafish danio rerio, a minnowlike freshwater fish, has emerged as a popular animal model for research and development. 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. Protocols are organized into sections corresponding to the chapters of the zebrafish book. The increase in zebrafish genomic resources together with more. Here we characterize the morphology and function of inflammatory macrophages in zebrafish larvae. Requirement of the fusogenic micropeptide myomixer for.
Left panels indicate input of the two proteins in the sample, and right panels are rfp antibody immunoprecipitated. View detailed rfp antibody specifications by linking to the specific product blocks. The specificity of this gfp antibody was confirmed by using isl1gfp fish higashijima. Gfp antibody recognizes the green fluorescent protein gfp, a fluorophore in widespread use in biomedical research. Zebrafish protein an overview sciencedirect topics. Rocklands anti rfp antibody can be used to detect native rfp and rfp.
This antibody can be used to detect rfp by elisa sandwich or capture for the direct binding of antigen. The protocol wiki contains protocols from the zebrafish book. Green fluorescent protein gfp expression directed by the hgj4a insertion. While the general breeding and handling of zebrafish are detailed in the zebrafish book 15, this. Human orthologs of this gene implicated in several diseases, including meesmann corneal dystrophy. Rfp embryos were injected with standard control std or tnfr2 mos. Immunohistochemistry of wholemount and sectioned zebrafish tissue. The single section of the spinal cord of hgj4a embryo at 48 hpf, stained with antipax2 antibody magenta. Quantitation of number of drgs and migrated drgs are indicated in panel c. B the 3d larvae labeled with antiacetylated tubulin antibody brown, lateral view with anterior to the left. 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.
Fluorescent proteins such as discosoma red fluorescent protein dsred from sea anemone discosoma sp. Pdf generation of twocolor transgenic zebrafish using. The superior specificity and sensitivity of this antibody. The monoclonal antibody facility regenerates supernatants and provides them to the zirc upon request. The sarcomere organization in slow muscles of homozygous myomesin3 rfp and myomesin3 knockdown zebrafish embryos. Hardcopies of the 4th edition of the zebrafish book can be obtained for a nominal fee from the zebrafish. Select rfp antibodies from monoclonal antibodies listed below. Regulatory elements that direct the glialspecific expression of zebrafish gfap were identified in genomic sequences from the zebrafish gfap gene. Fixed embryos can be kept in the fridge, but we noticed a loss of gfp signal.
Zebrafish protein antibody scbt santa cruz biotechnology. Generation of twocolor transgenic zebrafish using the green and red fluorescent protein reporter genes gfp and rfp article pdf available in marine biotechnology 42. This antibody also detects other gfp variants, including egfp and yfp, but not red fluorescent protein rfp. As the leading manufacturer of zebrafish antibodies, genetex offers an extensive portfolio of reagents validated for use in this model system. Rfp is a 53 kda ubiquitin proteasome system protein. A nonproteincoding gene is tagged and named novel transcript whereas a pseudogene. Similar refinements in the zebrafish data returned 1248 proteins with 2344 ppis in the. Rfp larvae stained at 3 dpf with antirfp keratinocytes, blue and antiphosphorylated h3 ph 3, proliferation marker antibodies. I have a transgenic mouse with a tdtomatotagged protein, so now i would like to immunoprecipitate the tdtomato and everything that is in the. Crisprz a database of crisprcas9 target sequences that have been experimentally validated in zebrafish. Description it has been 25 years since the now classic lab manual antibodies, by harlow and lane appeared. Rfp, also known as makorin ring finger protein 1 or rnf61, is another synonym for makorin ring finger protein 1. A guide for the laboratory use of zebrafish danio rerio. Reynolds is the first graphic novel from the popular fablevision publisher.
The formation of skeletal muscle fibers during embryogenesis and adult injuryinduced muscle repair occurs through the fusion of myoblasts. Left panels indicate input of the two proteins in the sample, and right panels are rfp antibody immunoprecipitated samples followed by western blot. Biotin conjugated polyclonal antirfp used in a sandwich elisa with unconjugated antirfp is well suited to titrate rfp. This second edition of antibodies, edited by edward a. The primordium that generates the embryonic posterior lateral line of zebrafish migrates from the head to the tip of the tail along a trail of sdf1producing cells.
One for alla highly efficient and versatile method for fluorescent. 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. Zebrafish have emerged as a powerful model system to study leukocyte recruitment and inflammation. Immunofluorescent analysis of hek293 cells transfected with pfin ef1mcherry vector labeling mcherry with ab167453 at 1500 red. Polyclonal anti rfp is designed detect rfp and its variants. 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. These cells are detected by the expression of the red fluorescent protein rfp. Select zebrafish protein antibodies from monoclonal antibodies listed below. 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. Formation of the spinal network in zebrafish determined by domain. Antibody data can be retrieved using the new antibody search form. The epithelialtomesenchymal transition emt is a highly conserved morphogenetic program essential for embryogenesis, regeneration and cancer metastasis. The prices we charge for products and services are to offset some of the actual costs we incur.
Green antibody staining is only seen in cells which express mcherry, as. Cells were fixed with formaldehyde, permeabilized using 0. Biotin conjugated polyclonal antirfp used in a sandwich elisa with unconjugated antirfp is well suited to titrate rfp in solution. The search form provides options to find antibodies that label specific anatomical structures or that target products of a specific gene. The monoclonal antibodies were generated with balbc mice against adult zebrafish. The zebrafish book, a guide for the laboratory use of zebrafish danio rerio. Methodologyprincipal finding here we present a novel method to. This is where zebrafish researchers can share experimental protocols and tips with the rest of the research community. A and wild type b embryos using antigfp antibodies abcam, ab290. Anti rfp tag antibody products red fluorescent protein, also known as rfp or dsred, is a 28kda fluorescent protein originating from the discosoma coral. The zebrafish danio rerio is one of these impressive cases, being able to regrow a fully functional and anatomically similar organ through epimorphic regeneration morgan, 1901. Gfp fluorescence allows realtime study of physiological processes involving fusion proteins or gfpexpressing cells. Designed as a part of a larger commitment to the childrens hospital boston, the book. Rfp proteins are useful markers for imagining protein localization, monitoring physiological processes, and detecting transgenic expression.
Functional analysis of slow myosin heavy chain 1 and. A representative frontal xy and lateral yz tridimensional reconstructions from confocal microscopy images of wihc of krt18. Due to their striking genomic similarity to humans, regenerative abilities, and shortened life cycle, zebrafish. View detailed zebrafish protein antibody specifications by linking to the specific product blocks. B zn5 wholemount immunostaining with ath5gap43rfp zebrafish transgenic line. A novel transgenic zebrafish model for bloodbrain and bloodretinal.
624 1251 938 63 1054 1209 660 786 814 310 2 1032 494 819 301 1358 576 1409 253 798 146 1474 112 1154 443 782 791 488 1520 558 69 1240 1143 673 195 208 755 959 1474 1437 1113 332 1236 970 877 1229 806 1403 1138