Anti-Human/Mouse HGAL - Dylight® 550 (H219-GRP)

HGAL (human germinal center-associated lymphoma) is a prognostic marker that predicts improved survival in patients with diffuse large B-cell or classic Hodgkin lymphoma1, 2, 3. HGAL protein is involved in several signaling pathways and cellular functions. HGAL decreases cell motility by interacting with F-actin and myosin II, inhibits the ability of myosin to translocate actin, and directly interacts with RhoA-specific guanine nucleotide exchange factors PDZ-RhoGEF and LARG4, 5, 6. HGAL also enhances intracellular B-cell receptor signaling by directly binding to Syk and enhancing Syk kinase activity7. Additionally, HGAL expression is upregulated after IL-4 stimulation8. HGAL protein localizes to both the cell cytoplasm and membrane2, 4, 7, 9, with a major proportion localizing to lipid rafts9. Localization is modulated by BCR activation and is facilitated by myristoylation and palmitoylation protein modifications9. Additionally, IL-6 induces HGAL phosphorylation and relocalization to podosome-like structures4. The anti-HGAL 1H1-A7 antibody was generated by immunizing mice with a GST-HGAL fusion protein and then fusing draining lymph node or spleen cells to K6H6B5 fusion partner hybridoma cells2. Hybridoma supernatants were screened by ELISA, subcloned, and then further screened by immunoblotting cellular lysates from stably transfected Daudi and HeLa cells expressing HGAL. 1H1-A7 is an IgG2a antibody containing a κ light chain. Human HGAL and its homologous mouse protein M17 are highly conserved but have some differences in expression pattern8. A comparison of real-time PCR and Western blotting data show a strong correlation between HGAL mRNA expression and protein abundance2.