The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 49 kDa (predicted molecular weight: 49 kDa).
Defects in GLA are the cause of Fabry disease (FD) [MIM:301500]. FD is a rare X-linked sphingolipidosis disease where glycolipid accumulates in many tissues. The disease consists of an inborn error of glycosphingolipid catabolism. FD patients show systemic accumulation of globotriaoslyceramide (Gb3) and related glycosphingolipids in the plasma and cellular lysosomes throughout the body. Clinical recognition in males results from characteristic skin lesions (angiokeratomas) over the lower trunk. Patients may show ocular deposits, febrile episodes, and burning pain in the extremities. Death results from renal failure, cardiac or cerebral complications of hypertension or other vascular disease. Heterozygous females may exhibit the disorder in an attenuated form, they are more likely to show corneal opacities.
Western blot - Galactosidase alpha antibody (ab70520)
Anti-Galactosidase alpha antibody (ab70520) at 1 µg/ml + Human liver tissue lysate - total protein (ab29889) at 10 µg
Secondary Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 49 kDa Observed band size : 49 kDa Additional bands at : 47 kDa (possible cleavage fragment).
Galactosidase alpha contains a 31 amino acid signal peptide. We hypothesize that the 47 kDa band represents the mature protein. Some batches of this product also recognise a band at 100 kDa in Western Blot, we are unsure of the identity of this band.
Citro V et al. Identification of an Allosteric Binding Site on Human Lysosomal Alpha-Galactosidase Opens the Way to New Pharmacological Chaperones for Fabry Disease. PLoS One11:e0165463 (2016).
Read more (PubMed: 27788225) »
Andreotti G et al. A thermodynamic assay to test pharmacological chaperones for Fabry disease. Biochim Biophys Acta1840:1214-24 (2014).
Read more (PubMed: 24361605) »