Glycogen Synthase Kinase 3 (GSK3) (FITC) (Drosophila)

Glycogen synthase kinase-3 (GSK-3), a multifunctional serine/threonine kinase, is a key regulator of numerous signaling pathways. Two isoforms of GSK-3 are reported in mammals: a 51kD GSK-3a and a 47kD GSK-3b. These two isoforms exhibit about 98% homology in their kinase domains, but share only about 36% identity in the last 76 C-terminal amino acid residues. GSK-3a contains a glycine-rich extension at its N-terminus. A minor (~15% of total) splice variant of GSK-3b, GSK-3b2, has also been identified, which contains a 13-residue insert within the kinase domain. It exhibits reduced kinase activity towards tau protein compared with ‘unspliced’ GSK-3b. GSK-3b2 is localized primarily to neuronal cell bodies, unlike unspliced GSK-3b that is also found in neuronal processes. GSK-3b plays a key inhibitory role in the Wnt signaling pathway. Wnt genes encode a large family of secreted, cysteine-rich proteins that are important in development and in maintenance of adult tissues. Abnormalities in Wnt signaling are reported to promote both human dgenerative diseases and cancer.

Applications:
Suitable for use in Western Blot and Immunocytochemistry. Not suitable for use in Immunoprecipitiation. Other applications not tested.

Recommended Dilution:
Western Blot: 0.5-2ug/ml. Detects GSK-3 in RIPA lysates from human Jurkat and A431, rat L6 and PC12 and mouse 3T3 cells. Jurkat cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with G8170-40 (0.5ug/ml). Proteins were visualized using an IgG (HRP) goat anti-mouse and a chemiluminescence detection system.
Immunocytochemistry: 10ug/ml. Demonstrated positive immunostaining for GSK3 in L6 cells fixed with acetone.
Optimal dilutions to be determined by the researcher.

Positive Control (Included): J9000: Jurkat Cell

Storage and Stability:
Store product at 4°C if to be used immediately within two weeks. For long-term storage, aliquot to avoid repeated freezing and thawing and store at -20°C. Aliquots are stable at -20°C for 12 months after receipt. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer.
Caution: FITC conjugates are sensitive to light. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.    

Note: Applications are based on unconjugated antibody.