Histone h2ax antibody staining protocol for immunohistochemistry. The specificity of this phosphospecific antibody was confirmed in several ways, showing that this antibody recognized neither the ectopically expressed s121a mutant nor endogenous mitotic h2ax. Due to their striking genomic similarity to humans, regenerative abilities. Nb100384 has been validated for use in human and mouse, it has not been tested nor validated for use in. We evaluated its possible genotoxic effect and oxidative stress in retinal cells of adult zebrafish exposed to 0. H2ax is abundant, fast, and correlates well with each dsb, it is the most sensitive marker that can be used to examine the dna damage produced and the. Serine 9 within this motif becomes rapidly phosphorylated to yield a form known as. 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 as model organism for drug discovery and toxicity testing.
Zebrafish antibodies primary antibodies sigmaaldrich. X phosphorylated ser9 antibody for flow cytometry 1. Past studies have also explored the ability of zebrafish to regenerate limbs and even hearts. H2ax is abundant, fast, and correlates well with each dsb, it is the most sensitive marker that can be used to examine the dna damage produced and the subsequent repair of the dna lesion. The phospho h2ax antibody was made against a phosphopeptide corresponding to residues 1142 of zebrafish histone h2ax. Expression of the histone h4 mutants in zebrafish embryos recapitulates. Zebrafish phosphorylated histone h2ax ph2ax was detected using a polyclonal antiph2ax antibody 1. Pdf zebrafish have a competent p53dependent nucleotide. The original zebrafish or zebra danio, danio rerio is a native of rivers in india and bangladesh. Cb2 is a reported alias name for the human gene cnr2, or cannabinoid receptor 2. Histone h2ax is a 14 kda ubiquitous member of the h2a histone family that contains an evolutionary conserved sq motif at the cterminus in eukaryotes. The zebrafish offers many advantages over other model systems thisse and zon, 2002, including ease of manual experimentation and drug administration and its prolific fecundity see table 1. It is available in convenient single piece packs or in cases of 25 each.
In rpdeficient zebrafish, we found activation of both nucleotide catabolism and. Jmse free fulltext toxicity of deltamethrin to zebrafish. Zebrafish are remarkably biologically similar to people and share the majority of the same genes as humans, making them an important model for understanding how genes work in health and. The zebrafish book a guide for the laboratory use of zebrafish danio brachydanio rerio by monte westerfield, institute of neuroscience, university of oregon older literature referred to zebrafish as.
I know this isnt large enough though of course, i didnt know at the time i purchased the fish, but i dont have the option of getting, say, a ten gallon tank. Our product portfolio covers all common host organisms and reactivity types. H2ax, which is deposited initially at the sites of dna doublestrand. Please use this form to recommend updates to the information in zfin. Zebrafish genome found strikingly similar to humans. A specific band was detected for histone h2ax at approximately 20 kda as indicated. Western blot analysis of extracts from 293 cells, untreated or uvtreated, using phosphohistone h2a. Manufactured from food grade polycarbonate virtually indestructible thereby minimizing replacement costs. Mefs were treated with uva 40 kjm 2 and parallel groups of h2axwt cells were treated with sp600125 or zvad for 1 h before uva radiation. Compare anti zebrafishfish antibody products from leading suppliers on biocompare. The hybridomas are stored at the monoclonal antibody facility of the uo under the supervision of dr. Deltamethrin is responsible for health and reproduction problems both in mammals and aquatic organisms. The cells were disrupted by adding dna stat60 teltest, inc.
About these antibodies all antibodies listed on this page were generated by researchers at the institute of neuroscience, university of oregon liu et al. Rabbit polyclonal antigamma h2ax p ser9 antibody dna doublestrand break marker cited in 90 publications. Rbbp4 is enriched in zebrafish neural stem and precursor cells during. Discard the supernatant and vortex to loosen cell pellet. In this study, zebrafish adults were exposed for 15 days to 0. Immunogen a portion of amino acids 258284 from the zebrafish protein was used as the immunogen for this pax2a antibody. Each kit contains zebrafish vtg, pre coated plates. H2ax showed that mst1 phosphorylates h2ax at ser9 fig.
Apoptosis is a fundamental process for metazoan development. Anticb2 antibody products listed below are anticb2 antibodies from multiple suppliers. A portion of amino acids 258284 from the zebrafish protein was used as the. Oct 01, 20 oocytespecific histone variants have been expected to play significant roles in early embryonic development, but the exact evidence and the biological function have remained unclear. Zebrafish have functional ifn proteins, as demonstrated by their protective effect on zebrafish cell lines infected with a fish rhabdovirus altmann et al. These foci, visible by immunofluorescence microscopy, occur at low frequency in cycling cells, reflecting atmdependent h2ax phosphorylation that occurs in mitosis ichijima 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 clones. This is where zebrafish researchers can help each other by sharing antibody information new antibodies, protocols, tips anything that might. 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. Zebrafish danio rerio, a minnowlike freshwater fish, has emerged as a popular animal model for research and development. I have three zebra danios in a one gallon fish tank. H2ax, made it possible to detect h2ax phosphorylation and thus detection of dna damage and repair in situ in individual cells. The zebrafish book a guide for the laboratory use of zebrafish danio brachydanio rerio by monte westerfield, institute of neuroscience, university of oregon older literature referred to zebrafish as brachydanio rerio.
Here, we present evidence that h2af1o, an oocytespecific h2a variant, is required for cell synchrony before midblastula transition in early zebrafish embryos. A guide for the laboratory use of zebrafish danio rerio. This is where zebrafish researchers can help each other by sharing antibody information new antibodies, protocols, tips anything that might be helpful. Mst1 promotes apoptosis through phosphorylation of histone h2ax. Oocytespecific h2a variant h2af1o is required for cell. Wb, simple western, flow, iccif, ihc, ihcfr, ihcp, chip, ko. The use of zebrafish to understand immunity sciencedirect. H2ax can be detected by immunoblotting and immunostaining using microscopic or flow cytometric detection. X ser9 antibody, clone jbw301, alexa fluor 647 emd millipore catalog. Apr 25, 20 zebrafish are remarkably biologically similar to people and share the majority of the same genes as humans, making them an important model for understanding how genes work in health and disease.
Retinoblastoma binding protein 4 maintains cycling neural stem. While the general breeding and handling of zebrafish are detailed in the zebrafish book 15, this. Emi1 maintains genomic integrity during zebrafish embryogenesis. We appreciate as much detail as possible and references as appropriate. Ab1runx1 from the zfin antibody database other names, clone ids, catalog ids etc. B, c rbbp4 and proliferative marker pcna antibody labeling of transverse. X antibody epr2282023 chip grade bsa and azide free ab256544 description. Duracross easily and effectively optimizes the breeding of zebra fish. Oct 30, 2019 welcome to the zfin community antibody wiki. In eukaryotes, dna double strand breaks dsbs have been. The type ii bacterial crisprcas9 system is rapidly becoming popular for genomeengineering due to its simplicity, flexibility and high efficiency.
Zebrafish immune system controls regeneration of eye. The vtg zebrafish elisa kit is a double antibody immunometric sandwich elisa for analyzing vtg in whole body homogenates and plasma samples. Can we apply antibody gamma h2ax p ser9 antibody nb100384 0. Select zebrafish protein antibodies from monoclonal antibodies listed below. H2ax in chromatin can be detected shortly after induction of dsbs in the form of discrete nuclear foci. The zebrafish as an emerging model to study dna damage in. Essential role of autoactivation circuitry on aurora b. Antibodies, primary antibodies, zfish zebrafish, alphabetical listing of zebrafish antibodies. X h2ax is a member of the histone h2a family which is one of the four core histones making up the nucleosome core particle. Zebrafish danio rerio is a fresh water fish that inhabits rivers in india. Zebrafish, danio rerio, are a species of freshwater, tropical fish within the minnow family i. Antiserum was subtracted against the unphosphorylated peptide and then affinity purified against the phosphopeptide. The marketplace for antibodies, elisa kits, and other research reagents. Cryopreservation and in vitro fertilization ivf protocols.
Antibodies research products elisa kits antibodies. Confocal microscopic images of hela cells, uv treated a and untreated b, showing nuclear stain with. Sgkkgssqsqey, where the phosphoserine is in brackets. Involved in cellular response to dna damage stimulus. The specificity of this phosphospecific antibody was confirmed in several ways, showing that this antibody recognized neither the ectopically expressed s121a mutant nor endogenous mitotic. To further confirm that mst1 phosphorylates h2ax at ser. View detailed zebrafish protein antibody specifications by linking to the specific product blocks. At the subcellular level, zebrafish and trout cells respond to various infectious stimuli similarly to mammals, suggesting that the interferoneif2. Western blot analysis of phosphohistone h2ax ser9 in a nih3t3 whole cell lysate untreated, b nih3t3 whole cell lysate 30m cisplatin treatment for 24hr using 30 g of protein. Maternal transfer and protective role of antibodies in. Actin dynamics 1 adherens junction dynamics 16 alzheimers disease 12 ampk signaling 10 angiogenesis 44 apoptosis regulation 44 b cell receptor signaling 47 death receptor signaling.
It is also relevant to the pathophysiology of immune diseases and cancers and to the outcome of cancer chemotherapies, as. Thaker department of veterinary pharmacology and toxicology, college. Select appropriate zebrafish protein antibodies for your research by isotype, epitope, applications and species reactivity. In this work, zebrafish embryos were obtained from control or. The role of the dna damage response in zebrafish and cellular. Duracross zebra fish breeding tank zebrafish breeding. Antibody level was measured by using indirect enzymelinked immunosorbent assay elisa method in the broodstock, egg, and fry. Danio rerio biology open the company of biologists. The vtg zebrafish elisa kit is a doubleantibody immunometric sandwich elisa for analyzing vtg in whole body homogenates and plasma samples. Both the histological and immunofluorescence if techniques showed the presence of apoptotic cells in the zebrafish retina after 9 days of treatment with 0.
It measures three centimeters long and has gold and dark blue stripes. I know this isnt large enough though of course, i didnt know at the time i purchased the fish, but i dont have the option of getting, say, a ten gallon tank because i cant take a big tank back and forth between my house and my school apartment its just not feasable. Oocytespecific histone variants have been expected to play significant roles in early embryonic development, but the exact evidence and the biological function have remained unclear. We investigated the chronic effects of deltamethrin on gonads by histopathological examination. Mouse monoclonal antigamma h2ax p ser9 antibody 9f3. Zebrafish protein antibody scbt santa cruz biotechnology. Zebrafish have a competent p53dependent nucleotide. To further confirm that mst1 phosphorylates h2ax at ser9, purified wildtype and mutant gsth2ax h2axs9a were used as substrates for an in vitro kinase assay. Cypermethrin cyp, is one of the most common contaminants in freshwater aquatic systems. Novel insights into the genetic and epigenetic paternal contribution to the human embryo.
Zebrafish types zebrafish aquaculture core facility at. Titration of the pax2a antibody may be required due to differences in protocols and secondarysubstrate sensitivity. Zebrafish as model organism for drug discovery and. Together, these observations provide novel evidence that elevated expression of.
Mst1 promotes apoptosis through phosphorylation of histone. 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. Zebrafish immune system controls regeneration of eye tissue. H2ax foci are an early marker of dsb sites undergoing dna repair. Genotoxicity induced by cypermethrin in the zebrafish retina. We evaluated its possible genotoxic effect and oxidative stress in retinal cells of adult zebrafish exposed. In eukaryotes, dna double strand breaks dsbs have been shown to trigger the phosphorylation of serine 9 at the carboxy terminus of histone h2ax resulting in gammah2ax. Histone h2ax is a 14 kda ubiquitous member of the h2a histone family that contains an evolutionary conserved sq motif at.
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