To view the continuously updated ihcicc image gallery, please visit rndsystems. Ihc fr image of doublecortin staining in mouse adult dentate gyrus sections using ab187231. Doublecortin is a microtubule associated protein that stabilizes and bundles microtubules. The mission of the national institute of environmental health sciences is to discover how the environment affects people in order to promote healthier lives. The neuroanatomical distribution of doublecortin expression does not match the pattern of neurogenesis in the canary brain. Immunohistochemistry technically, ihc and icc are relatively simple and. Do u b le c o r tin an tib o d y cell signaling technology. Ab2253, is a highly specific guinea pig polyclonal antibody that targets doublecortin and has been tested for use in immunocytochemistry. Brain and behavior research foundation the robert n.
Please optimize the procedure since experimental conditions can vary for. Immunohistochemistry instruction manual immunostar, inc. Brdu is incorporated into replicating dna and can be detected using antibrdu antibodies. Vellema and colleagues report that dcxexpressing cells, based both on immunohistochemistry for the protein and in situ hybridization for the corresponding mrna, are found throughout the canary brain, with the most pronounced expression in the telencephalon.
Doublecortin antibody lsc36204 is an unconjugated rabbit polyclonal antibody to doublecortin dcx from human. A conserved doublecortin domain mediates the interaction with microtubules, and interestingly most missense. Antidoublecortin antibody antidoublecortin antibody, cat. We incorporated these steps into a 3d antibody staining protocol for the adult. The sections were fixed in paraformaldehyde and permeabilized using 1x tbst. Incubate with primary antibody goat polyclonal antidcx 1. Datasheet for goat antidoublecortin dcx aa232242 antibody eb10781 a polyclonal antibody designed to detect protein products of the gene dcx by western blot and elisa. Immunohistochemistry formalinpfafixed paraffinembedded sections anti doublecortin antibody 2g5 ab5349 immunohistochemical analysis of paraffinembedded human kidney cancer tissue labelling doublecortin with ab5349 at 1200 dilution followed by dab staining. The aim of the present immunohistochemical study was to evaluate the expression of these proteins and their pattern of distribution in canine brain, including agerelated changes, and in nonnervous tissues. Chose your productspecific protocol pdf from a range of antibody applications. Dehydrate the tissue through 70%, 80%, 95% alcohol, 5 min each. Box 488 hudson, wisconsin 540160488 tollfree phone. Immunohistochemistry and multiple labeling with antibodies from.
Mutations in the human dcx gene cause lissencephaly. The experimental protocol was approved by the institutional animal care and use committee iacuc of seoul national university, south korea approval no. Immunohistochemistry ihc is a technique used to analyze protein expression in the context of tissue morphology. Fix freshly dissected tissue ihcicc protocol guide 4 tissue and cell samples must be appropriately harvested and prepared for each ihcicc study. Doublecortin immunohistochemistry protocol the denny. Primary article validation of doublecortin hippocampal.
Immunohistochemistry ihc uses antibodies to detect the location of proteins and other antigens in tissue sections. Doublecortin, however, is not an exclusive marker of neural stem cells, but also of neuronal precursors. The sections were then blocked using 10% donkey serum for 1 hour at 25c. Immunohistochemistry protocol for paraffinembedded tissue. Goat antidoublecortin dcx aa232242 antibody eb10781. Immunohistochemical study of doublecortin and nucleostemin in. Immunohistochemical study of doublecortin and nucleostemin. Properties of doublecortindcxexpressing cells in the.
Histology and immunohistochemistry protocols from other online sources. Doublecortin antibody lsc204512 is an unconjugated mouse monoclonal antibody to doublecortin dcx from human, mouse, rat and other species. The results confirmed that the immunohistochemistry protocol supplied by denny et al. The antibodyantigen interaction is visualized using either chromogenic detection. Prepare formalinfixed, paraffinembedded tissue sections step 18. Doublecortin is a highly valuable endogenous marker of. The piriform cortex receives input from the olfactory bulb and via the entorhinal cortex sends efferents to the hippocampus, thereby connecting the two canonical neurogenic regions of the adult rodent. A conserved doublecortin domain mediates the interaction with microtubules, and interestingly most missense mutations cluster in this domain 2. Please refer to the western blot protocol found on the product web. Frozen section technique a method for preparation of frozen. Tested in western blot wb, immunofluorescence if, immunocytochemistry icc and immunoprecipitation ip applications.
Colabeling of dcx together with neun, which is expressed in postmitotic neurons, allows the. Brains stained for glial fibrillary acidic protein, doublecortin and. Arpita kabiraj, jagriti gupta, tanya khaitan, preeti tomar bhattacharya int j biol med res. Kinases jnk, cdk5 and pka phosphorylate doublecortin. Immunohistochemistry protocol for frozen tissue download a pdf. Place the sterile cover slips in 12 or 24 well plates. Immunohistochemistry ihc uses antibodies to detect cell and tissue proteins and provide semiquantitative data about target protein expression, distribution, and localization. Read more about protocol for the preparation and fluorescent ihc staining of frozen tissue sections troubleshooting guide.
This video article illustrates a comprehensive protocol to detect and quantify all stages of adult. To facilitate the required incubation steps, whole tissues must be cut into ultra thin. Plate the cells on the cover slips at a density of 10,000. Protocol for immunohistochemistry freefloating sections with antidoublecortin antibody for avian tissue. A method for 3d immunostaining and optical imaging of the mouse. Histology and immunohistochemistry online protocols. Alternatively, you can find product specific protocols in the protocols tab on every antibody product page. Protocol for the preparation and fluorescent ihc staining. Vellema and colleagues report that dcxexpressing cells, based both.
Immunohistochemistry ihc combines anatomical, immunological and biochemical techniques to image discrete components in tissues by using appropriatelylabeled antibodies to bind specifically to their. It was reported that the proteins doublecortin and nucleostemin were related with stemprogenitor cells in central nervous system. Each antibody has its own optimum fixation conditions, see protocol 5 for a discussion of how to determine those conditions. The researcher should optimize the precise conditions for a particular assay. Wash slides with specific reagents in the following order. For staining in wild type versus transgenic strains. This article describes an ihc protocol optimized by scientists at cell signaling. Immunohistochemistry formalinpfafixed paraffinembedded sections antidoublecortin antibody ab18723 ihc image of ab18723 staining in mouse 8 weeks brain formalin fixed paraffin embedded. Columbia university medical center new york state psychiatric institute research foundation for mental hygiene, inc. Immunohistochemistry formalinpfafixed paraffinembedded sections antidoublecortin antibody epr19997 ab207175 ihc image of doublecortin staining in a formalin fixed, paraffin embedded. Principle and techniques of immunohistochemistry a. Validation of doublecortin hippocampal localization.
Validated in wb, ihc, icc, iccif and tested in mouse, rat, human, quail. In the following experiment, an immunohistochemistry protocol supplied by denny et al. Pdf potential adult neurogenesis in the telencephalon. Immunohistochemistry is a method of detecting the presence of specific proteins in cells or tissues and consists of the following steps.1303 1383 946 1495 1611 497 1496 497 63 1199 714 967 751 117 1407 1132 510 267 1615 1524 1120 1496 120 896 840 743 582 236 776 1346 1040