Anti-Wnt8b (Wingless-type MMTV (Mouse Mammary Tumor Virus) Integration Site Family Member)

Mouse Wnt-8b (Wingless related integration site family member 8b) is a 40-50 kD secreted glycoprotein member of the Wnt family of proteins that function as ligands for the Frizzled family of receptors. Mature mouse Wnt-8b is 329 aa in length and contains 24 cysteines plus three potential N-linked glycosylation sites. Little is known about Wnt-8b. It appears to play a role in forebrain development. The mature mouse protein is 100%, 98% and 97% aa identical to mature rat, canine and human Wnt-8b, respectively. Hybridoma: Produced from a hybridoma resulting from the fusion of a mouse myeloma with B cells obtained from a rat immunized with a purified, E.coli-derived, recombinant protein consisting of mouse Wnt-8b amino acid (aa) segments 22-52 and 195-269 (Accession # AAB31816) fused via a gly/ser linker (rmWnt-8b). Applications: Western Blot: This antibody can be used at 1-2ug/ml with the appropriate secondary reagents to detect human or mouse Wnt-8b. Using a colorimetric detection system, the detection limit for rmWnt-8b is approximately 25 ng/lane under non-reducing and reducing conditions. Chemiluminescent detection will increase sensitivity by 5 to 50 fold. This antibody was used at 5ug/ml to detect Wnt-8b in cell extracts. Western Blot buffers are listed below. Wash Buffer Blocking Buffer Blotting Buffer 25 mM Tris, pH 7.4 5% nonfat dry milk 2% nonfat dry milk, 0.5 M NaCl in Wash Buffer in Wash Buffer 0.05% Tween(R) 20 , Direct ELISA: This antibody can be used at 0.5-1.0ug/ml with the appropriate secondary reagents to detect human or mouse Wnt-8b. The detection limit for rmWnt-8b is approximately 5 ng/well. , Optimal dilutions should be determined by each laboratory for each application. Storage and Stability: Lyophilized powder may be stored at 4°C for short-term only. Reconstitute to nominal volume by adding sterile PBS with 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.