| Solution Information | help | |
| Enzyme: | Ephrin type-A receptor 4 | |
| inhibitor: | BDBM44502 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | EphA4 assay materials: 1) EphA4-AlkalinePhosphatase conditioned media (from EphA4-AP secreting stable cell-line) (cell line: NIH3T3 stably transfected with EphA4-AP), as well as control Alkaline Phosphatase conditioned media, was provided by Dr. Elena Pasquale (Sanford-Burnham Medical Research Institute, San Diego, CA). 2) KYL-biotinylated peptide (KYLPYWPVLSSL), re-constituted in PBS, was provided by Dr. Elena Pasquale. 3) Streptavidin (Pierce Cat # PI21125) 1mg/ml in 0.1M Boric Acid pH 8.7 4) Blocking buffer: 5mg/ml BSA in 20mM Tris pH 7.5, 150mM NaCl 5) Assay/Washing buffer: 20mM Tris pH 7.5, 150mM NaCl, 1mM CaCl2, 0.01% Tween-20 6) Detection reagent: 1mg/ml pNPP in 1.1M Diethanolamine, 0.5mM MgCl2 pH 9.8 7) Stop Reagent: 1N NaOH EphA4 binding assay protocol: Day 1 1) Add 25ul of streptavidin (1ug/ml) to each well of a microtiter plate 2) Incubate, at least overnight (18hrs), at room temp (keep plates covered and in the dark) *after the overnight incubation, plates can be place | |
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