Human Doublecortin ELISA Kit (ab218267)
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- References
- Protocols
Overview
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Product nameHuman Doublecortin ELISA Kit
See all Doublecortin kits -
Detection methodColorimetric
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Precision
Intra-assay Sample n Mean SD CV% Overall 3 4.2% Inter-assay Sample n Mean SD CV% Overall 5 5.8% -
Sample typeCell culture extracts, Tissue Extracts
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Assay typeSandwich (quantitative)
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Sensitivity2.13 pg/ml
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Range15.63 pg/ml - 1000 pg/ml
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Recovery
Sample specific recovery Sample type Average % Range Cell culture extracts 105 97% - 118% Tissue Extracts 111 98% - 120% Mouse brain extract 96 89% - 106% -
Assay time1h 30m
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Assay durationOne step assay
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Species reactivityReacts with: Mouse, Rat, Human
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Product overview
Doublecortin in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Doublecortin protein in human cell and tissue extract samples.
The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.
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Notes
Doublecortin is a microtubule-associated protein required for initial steps of neuronal dispersion and cortex lamination during cerebral cortex development. It may act by competing with the putative neuronal protein kinase DCLK1 in binding to a target protein. Doublecortin may in that way participate in a signaling pathway that is crucial for neuronal interaction before and during migration, possibly as part of a calcium ion-dependent signal transduction pathway. Doublecortin may be part with PAFAH1B1/LIS-1 of overlapping, but distinct, signaling pathways that promote neuronal migration. Doublecortin interacts with tubulin and USP9X.
Mutations affecting the Doublecortin gene are causing X-linked 1 Lissencephaly, a disease characterized by mental retardation and seizures that are more severe in male patients. Female patients display a less severe phenotype referred to as 'doublecortex'.
Doublecortin is highly expressed in neuronal cells of fetal brain (in the majority of cells of the cortical plate, intermediate zone and ventricular zone), but not expressed in other fetal tissues. In the adult, it is highly expressed in the brain frontal lobe, but it has very low expression in other regions of brain, and it is not detected in heart, placenta, lung, liver, skeletal muscles, kidney and pancreas.
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Tested applicationsSuitable for: Sandwich ELISAmore details
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PlatformPre-coated microplate (12 x 8 well strips)
Properties
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Storage instructionsStore at +4°C. Please refer to protocols.
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Components 1 x 96 tests Human Doublecortin Lyophilized Recombinant Protein 2 vials 10X Human Doublecortin Capture Antibody 1 x 600µl 10X Human Doublecortin Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent CPI 1 x 6ml Plate Seals 1 unit Sample Diluent NS 1 x 12ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml -
Research areas
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FunctionSeems to be required for initial steps of neuronal dispersion and cortex lamination during cerebral cortex development. May act by competing with the putative neuronal protein kinase DCAMKL1 in binding to a target protein. May in that way participate in a signaling pathway that is crucial for neuronal interaction before and during migration, possibly as part of a calcium ion-dependent signal transduction pathway. May be part with LIS-1 of an overlapping, but distinct, signaling pathways that promote neuronal migration.
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Tissue specificityHighly expressed in neuronal cells of fetal brain (in the majority of cells of the cortical plate, intermediate zone and ventricular zone), but not expressed in other fetal tissues. In the adult, highly expressed in the brain frontal lobe, but very low expression in other regions of brain, and not detected in heart, placenta, lung, liver, skeletal muscles, kidney and pancreas.
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Involvement in diseaseDefects in DCX are the cause of lissencephaly X-linked type 1 (LISX1) [MIM:300067]; also called X-LIS or LIS. LISX1 is a classic lissencephaly characterized by mental retardation and seizures that are more severe in male patients. Affected boys show an abnormally thick cortex with absent or severely reduced gyri. Clinical manifestations include feeding problems, abnormal muscular tone, seizures and severe to profound psychomotor retardation. Female patients display a less severe phenotype referred to as 'doublecortex'.
Defects in DCX are the cause of subcortical band heterotopia X-linked (SBHX) [MIM:300067]; also known as double cortex or subcortical laminar heterotopia (SCLH). SBHX is a mild brain malformation of the lissencephaly spectrum. It is characterized by bilateral and symmetric plates or bands of gray matter found in the central white matter between the cortex and cerebral ventricles, cerebral convolutions usually appearing normal.
Note=A chromosomal aberration involving DCX is found in lissencephaly. Translocation t(X;2)(q22.3;p25.1). -
Sequence similaritiesContains 2 doublecortin domains.
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Cellular localizationCytoplasm.
- Information by UniProt
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Alternative names
- DBCN
- Dbct
- DC
see all -
Database links
- Entrez Gene: 1641 Human
- Entrez Gene: 13193 Mouse
- Entrez Gene: 84394 Rat
- Omim: 300121 Human
- SwissProt: O43602 Human
- SwissProt: O88809 Mouse
- SwissProt: Q9ESI7 Rat
- Unigene: 34780 Human
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Associated products
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Alternative Versions
Applications
Our Abpromise guarantee covers the use of ab218267 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| Sandwich ELISA | Use at an assay dependent concentration. |
Images
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Example of human Doublecortin standard curveBackground-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of native Doublecortin in SH-SY5Y cell extract, human fetal brain tissue extract and rat brain tissue extract.Interpolated concentrations of native Doublecortin in SH-SY5Y cell extract samples based on a 100 µg/mL extract load, human fetal brain tissue extract samples based on a 300 µg/mL extract load, and rat brain tissue extract samples based on a 500 µg/mL extract load. The concentrations of Doublecortin were measured in duplicate and interpolated from the Doublecortin standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Doublecortin concentration was determined to be 703 pg/mL in SH-SY5Y cell extract samples, 415 pg/mL in human fetal brain tissue extract samples, and 535 pg/mL in rat brain tissue extract samples.
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Doublecortin is highly expressed in neuronal cells of the fetal brain. Human adult brain tissue extract and human fetal brain tissue extract samples were analyzed based on a 300 µg/mL extract load.The concentrations of Doublecortin were measured in duplicate and interpolated from the Doublecortin standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted in pg Doublecortin per mg of extract (mean +/- SD, n=3). The mean Doublecortin concentration was determined to be 309 pg/mg in human adult brain tissue extract samples, and 1375 pg/mg in human fetal brain tissue extract samples.
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Other species reactivity was determined by measuring various species adult brain tissue extract samples.The concentrations of Doublecortin were measured in duplicate and interpolated from the Doublecortin standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted in pg Doublecortin per mg of extract (mean +/- SD, n=3). The mean Doublecortin concentration was determined to be 309 pg/mg in human adult brain tissue extract samples, 147 pg/mg in mouse brain tissue extract samples, and 878 pg/mg in rat brain tissue extract samples.
Protocols
References
ab218267 has not yet been referenced specifically in any publications.
