Western Blot: This antibody can be used at 0.1-0.2ug/ml with the appropriate secondary reagents to detect human HES-4. The detection limit for rhHES-4 is approximately 5 ng/lane under non-reducing and reducing conditions. Immunocytochemistry: This antibody has been used at a concentration of 10ug/ml to detect HES-4 in HEK293 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 fluorochrome-coupled anti-sheep 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 HES-4. The detection limit for rhHES-4 is approximately 0.2 ng/well. Optimal dilutions should be determined by each laboratory for each application.

Lyophilized and reconstituted products are stable for 12 months after receipt at -20°C. Reconstitute with sterile PBS. Aliquot to avoid repeated freezing and thawing. Store 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.

E. coli-derived rhHES-4 (aa94-221)

Supplied as a lyophilized powder in PBS, 5% trehalose. Reconstitute with 500ul sterile PBS.

Purified by immunoaffinity chromatography.

Selected for its ability to recognize human HES-4 in the applications listed below. In direct ELISAs and Western Blots, this antibody shows less than 1% crossreactivity with rhHES-1, rhHES-2, rhHES-5, and rhHES-7.