Western Blot: This antibody can be used at 0.1-0.2ug/ml with the appropriate secondary reagents to detect Wnt-2b. The detection limit for rmWnt-2b is approximately 5 ng/lane and 1 ng/lane under non-reducing and reducing conditions, respectively. In this format, this antibody shows approximately 20% cross-reactivity with rhWnt-2, and less than 5% cross-reactivity with rhWnt-7a, -9a, rmWnt-1, -3a, -4, -5a, -5b, -8a, -8b, -9b, -10a, -10b, -11, -16. Immunocytochemistry: This antibody has been used at a concentration of 10ug/ml to detect Wnt-2b in BT20 human breast carcinoma 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 NorthernLightsTM-557-coupled anti-goat IgG at room temperature in the dark for one hour. Between each step, cells were washed with PBS containing BSA. Direct ELISA: This antibody can be used at 0.5-1.0ug/ml with the appropriate secondary reagents to detect mouse Wnt-2b. The detection limit for rmWnt-2b is approximately 0.5 ng/well. Optimal dilutions should be determined by each laboratory for each application.

Reconstitute with sterile PBS. If 0.5 mL of PBS is used, the antibody concentration will be 0.2 mg/mL.

Lyophilized powder may be stored at 4°C for short-term only. Reconstitute to nominal volume by adding sterile 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.

E. coli-derived rmWnt-2b

Supplied as a lyophilized powder from a 0.2um filtered solution in PBS, 5% trehalose.

Purified by mouse Wnt-2b affinity chromatography.

Selected for its ability to recognize mouse and human Wnt-2b in the applications listed.