Anti-GLi3 (GLI-Kruppel Family Member GLI3)

Human GLI-3 is a 190kD transcription factor of the C2H2-type zinc-finger protein family. GLI-3 appears to play a role in limb and brain development, probably through sonic hedgehog signaling. GLI-3 is regulated by PKA-mediated phosphorylation. Unphosphorylated full-length GLI-3 is a transcriptional activator, while phosphorylated GLI-3 is cleaved after amino acid 700 to generate an N-terminal 83kD transcriptional repressor. Over the region used as immunogen, human GLI-3 is 95 % identical to the corresponding mouse and canine protein sequences. Applications: Suitable for use in ELISA, Immunocytochemistry, and Western Blot. Other applications have not been tested. Recommended Dilution: Western Blot: 0.1-0.2ug/ml. Used in conjunction with the appropriate secondary reagents, detects human and mouse GLI-3. The detection limit for rhGLI-3 and rmGLI-3 is approximately 5 ng/lane under non- reducing and reducing conditions. , Immunocytochemistry: 10ug/ml detects human GLI-3 in HeLa cells. Cells were fixed with PBS containing 4% paraformaldehyde and blocked with PBS containing 10% normal donkey serum, 0.1% Triton X-100, and 1% BSA. After blocking, cells were incubated with diluted primary antibody followed by NorthernLights 557-coupled anti-goat IgG at room temperature in the dark. Between each step, cells were washed with PBS containing BSA. , Direct ELISA: 0.5-1.0ug/ml. Used in conjunctionthe appropriate secondary reagents, detects human and mouse GLI-3. The detection limit for rhGLI-3 and rmGLI-3 is approximately 1 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 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.