Anti-Human/Mouse HGAL – Dylight® 650

This DyLight® 650 conjugate is formulated in 0.01 M phosphate buffered saline (150 mM NaCl) PBS pH 7.4, 1% BSA and 0.09% sodium azide as a preservative.

This DyLight® 650 conjugate is stable when stored at 2-8°C. Do not freeze.

Red Laser (652 nm)

IHC^{2, 7},
IP^{4, 6, 7, 9},
IF Microscopy⁴

1H1-A7 activity is directed against human and mouse HGAL.

HGAL is expressed in germinal center (GC) lymphocytes, spleen, thymus, and many lymphoma subtypes, predominantly in GC-derived lymphomas and particularly in B-cell and Hodgkin lymphomas.

HGAL (human germinal center-associated lymphoma) is a prognostic marker that predicts improved survival in patients with diffuse large B-cell or classic Hodgkin lymphoma^{1, 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 LARG^{4, 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 membrane^{2, 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.

1. Lossos IS, Alizadeh AA, Rajapaksa R, et al. Blood. 101(2):433-440. 2003.
2. Natkunam Y, Lossos IS, Taidi B, et al. Blood. 15;105(10):3979-3986. 2005.
3. Azambuja D, Lossos IS, Biasoli I, et al. Leuk Lymphoma. 50(11):1830–1836. 2009.
4. Lu X, Chen J, Malumbres R, et al. Blood. 110(13):4268-4277. 2007.
5. Jiang X, Lu X, McNamara G, et al. Blood. 116(24):5217–5227. 2010.
6. Lu X, Kazmierczak K, Jiang X, et al. FEBS J. 278(11):1922–1931. 2011.
7. Romero-Camarero I, Jiang X, Natkunam Y, et al. Nat Commun. 4:1338. 2013.
8. Christoph T, Rickert R, Rajewsky K. Int Immunol. 6(8):1203-11. 1994.
9. Lu X, Sicard R, Jiang X, et al. Blood. 125(4):649-57. 2015.