DNER (Delta and Notch-like Epidermal Growth Factor-related Receptor, Brain EGF Repeat Containing Transmembrane Protein, BET)

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.\n\nApplications: \nWestern 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. \nFlow 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 microL of the diluted solution to 1-2.5 x 10e5 cells in a total reaction volume not exceeding 200 microL. The binding of unlabeled antibodies may be visualized by adding 10 microL of a 25ug/ml stock solution of a secondary developing reagent such as anti-goat lgG conjugated to a fluorochrome. \nImmunocytochemistry: 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 degrees 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. \nDirect 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.\nOptimal dilutions should be determined by each laboratory for each application.\n\nStorage and Stability:\nLyophilized powder may be stored at 4 degrees C for short-term only. Reconstitute to nominal volume by adding sterile PBS with 40-50% glycerol and store at -20 degrees C. Reconstituted product is stable for 12 months at -20 degrees 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.

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SPECIFICATIONS

Catalog Number

D4016-85

Size

100ug

Applications

ELISA, FC, ICC, WB

Hosts

Goat

Reactivities

Hum

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.

P Type

Pab

Purity

Purified by human DNER affinity chromatography.

Isotype

IgG

Additional Info

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.