The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
WB: 1/500 sufficient for detection of a-Glucosidase II in 20µg of rat brain tissue extract by colorimetric analysis.
It is recommended for Western blot applications, that the secondary antibody be quenched by adding normal sera (of an identical species to secondary host) to the diluent. Detects a band of approximately 110 kDa.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Cleaves sequentially the 2 innermost alpha-1,3-linked glucose residues from the Glc(2)Man(9)GlcNAc(2) oligosaccharide precursor of immature glycoproteins.
Detected in placenta.
Glycan metabolism; N-glycan metabolism.
Belongs to the glycosyl hydrolase 31 family.
Endoplasmic reticulum. Golgi apparatus. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
Immunocytochemistry/ Immunofluorescence - Anti-alpha Glucosidase II antibody (ab69489)
ICC/IF image of ab69489 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab69489 at 1/200 dilution overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in methanol (100%, 5min) and formaldehyde (4%, 10min) fixed HepG2 cells at 1/200 dilution.
Iwamoto S et al. Reconstructed glycan profile for evaluation of operating status of the endoplasmic reticulum glycoprotein quality control. Glycobiology23:121-31 (2013).
Read more (PubMed: 22975980) »
Dai L et al. Differential profiling studies of N-linked glycoproteins in glioblastoma cancer stem cells upon treatment with ?-secretase inhibitor. Proteomics11:4021-8 (2011).
Read more (PubMed: 21898824) »