| DNER, also known as brain EGF repeat containing transmembrane protein (BET), is a neuron-specific type I transmembrane notch ligand that is expressed on membranes of dendrites and cell bodies but not axons. It contains ten extracellular EGF-like domains and one follistatin-like domain (between the 9th and 10th EGF-like domains). DNER mediates neuron-glia interactions and is required for the functional and morphological maturation of the cerebellum. The extracellular domains of human and mouse DNER share 89% amino acid sequence homology. |
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| Catalog # | D4016-85 |
| Applications | Western Blot: This antibody can be used at 0.1-0.2ug/ml with the appropriate secondary reagents to detect human DNER. The detection limit for rhDNER is approximately 1 ng/lane and 5 ng/lane under non-reducing and reducing conditions, respectively. |
| Flow Cytometry: This antibody has been tested on A172 human glioblastoma cells for use in flow cytometry. Dilute this antibody to 50ug/ml and add 10 L of the diluted solution to 1-2.5 x 10e5 cells in a total reaction volume not exceeding 200 L. The binding of unlabeled antibodies may be visualized by adding 10 L of a 25ug/ml stock solution of a secondary developing reagent such as anti-goat lgG conjugated to a fluorochrome. |
| Immunocytochemistry: This antibody has been used at a concentration of 10ug/ml to detect DNER in A172 human glioblastoma cells. Cells were fixed with PBS containing 4% paraformaldehyde for 20 minutes at room temperature and blocked with PBS containing 10% normal donkey serum, 0.1% Triton X-100, and 1% BSA for 45 minutes at room temperature. After blocking, cells were incubated with diluted primary antibody overnight at 4° C followed by Rhodamine RedTM-coupled anti-goat IgG at room temperature in the dark for one hour. Between each step, cells were washed with PBS containing 0.1% BSA. |
| Direct ELISA: This antibody can be used at 0.5-1.0ug/ml with the appropriate secondary reagents to detect human DNER. The detection limit for rhDNER is approximately 0.2 ng/well. |
| Optimal dilutions should be determined by each laboratory for each application. |
| Storage and Stability | Lyophilized powder may be stored at 4°C for short-term only. Reconstitute to nominal volume by adding sterile PBS with 40-50% glycerol and store at -20°C. Reconstituted product is stable for 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer. |
| Clone Type | Polyclonal |
| Isotype | IgG |
| Host | Goat |
| Source | Human |
| Concentration | As reported |
| Form | Supplied as a lyophilized powder from a 0.2um filtered solution in PBS with 5% trehalose. Reconstitute with sterile PBS, 40-50% glycerol. If 0.5ml is used, the concentration will be 0.2mg/ml. |
| Purity | Purified by human DNER affinity chromatography. |
| Immunogen | NS0-derived rhDNER extracellular domain |
| Specificity | Selected for its ability to recognize human DNER in the applications listed. In direct ELISAs and Western Blots, this antibody shows ~30% crossreactivity with rmDNER. |
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| Important Note | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. |
