Antibody DetailsProduct DetailsReactivity Species Rat Host Species Mouse Immunogen NS0-Derived Recombinant Rat EphA5 Extracellular Domain Endotoxin Level <0.1 EU/µg as determined by the LAL method Formulation This monoclonal antibody has been 0.2 µm filtered and lyophilized from modified Dulbecco's phosphate buffered saline (1X PBS) pH 7.2 - 7.3 containing 5.0% w/v trehalose with no calcium, magnesium or preservatives present. Storage and Handling The lyophilized antibody can be stored desiccated at -20°C to -70°C for up to twelve months. The reconstituted antibody can be stored for at least four weeks at 2-8°C. For long-term storage of the reconstituted antibody, aseptically aliquot into working volumes and store at -20°C to -70°C in a manual defrost freezer. Avoid repeated freeze thaw cycles. No detectable loss of activity was observed after six months. Country of Origin USA Shipping Next Day Ambient RRIDAB_2830026 Applications and Recommended Usage? Quality Tested by Leinco Flow Cytometry: It is recommended to use the indirect method for signal enhancement when enumerating cells expressing EphA5. A suggested method would be to stain cells expressing EphA5 with 0.25 µg per 1 - 2.5 x 106 cells in a total staining volume of ≤200 µl followed by PN:M1189. Western Blotting: To detect Rat EphA5 this monoclonal antibody can be used at a concentration of 1-2 µg/ml. This monoclonal antibody should be used in conjunction with compatible second-step reagents such as PN:M114 and a chromogenic substrate such as PN:T343. The detection limit for Rat EphA5 is 1 ng/lane under either reducing or non-reducing conditions. The sensitivity of detection may increase up to 50 fold when a chemiluminescent substrate is used. Other Applications Reported In Literature ? Immunocytochemistry: This antibody was used at a concentration of 10 μg/mL to detect EphA5 on U-118-MG 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 Red™-coupled anti-mouse IgG at room temperature in the dark for one hour. Between each step, cells were washed with PBS containing 0.1% BSA. CyTOF-ready: Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation. Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change. DescriptionSpecificity Mouse Anti-Rat EphA5 (Clone 86731) recognizes an epitope on Rat EphA5. This monoclonal antibody was purified using multi-step affinity chromatography methods such as Protein A or G depending on the species and isotype. No cross reactivity was detected with rhEphA1, rmEphA2, -A3, -A6, -A7, rrEphB1, -B2 or rmEphB3. Background EPH receptor A5 (EPHA5) is a glycoprotein that is expressed by astrocytes and skeletal muscle. It provides guidance cues for retinal ganglion cells1 and is essential for proper retinal axon guidance and topographic mapping in the mammalian visual system.2 Ephrin A5 serves as a general inhibitor of axonal growth from limbic neurons.3 It also plays a role in motor axon outgrowth and pathfinding to the hindlimb.4,5 Antigen DetailsProtein PubMed NCBI Gene Bank ID UniProt.org References & Citations1. Davenport, RW. et al. (1998) J Neurosci. 18: 975 2. Frisen, J. et al. (1998) Neuron 20: 235 3. Gao, PP. et al. (1998) Proceedings of the National Acad of Sci. 95: 5329 4. Eberhart, J. et al. (2000) Developmental Neurosci. 22: 237 5. Eberhart, J. et al. (2004) J of Neurosci. 24: 1070 Technical Protocols  |
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