1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Ubiquitous. Higher expression levels observed in the temporal lobe and fetal brain.
Defects in RNASET2 are the cause of leukoencephalopathy cystic without megalencephaly (LCWM) [MIM:612951]. An infantile-onset syndrome of cerebral leukoencephalopathy. Affected newborns develop microcephaly and neurologic abnormalities including psychomotor impairment, seizures and sensorineural hearing impairment. The brain shows multifocal white matter lesions, anterior temporal lobe subcortical cysts, pericystic abnormal myelination, ventriculomegaly and intracranial calcifications.
Belongs to the RNase T2 family.
Secreted. Subcellular fractionation of transfected ovarian cancer cells reveals full-length RNASET2 in the endoplasmic reticulum fraction and the 2 smaller RNASET2 proteolytic products in the lysosome fraction.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen carcinoma tissue sections labeling Ribonuclease T2 with ab107313 at 1/25 dilution. Tissue was fixed with formaldehyde and blocked with 3% BSA for 0.5 hour at 37°C. Heat mediated antigen retrieval was performed using a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hour at 37°C. A Peroxidase-conjugated Goat anti-rabbit polyclonal (ready to use) was used as the secondary antibody.
ab107313 stained LoVo 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 ab107313 at 5µg/ml 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.
Western blot - Ribonuclease T2 antibody (ab107313)
Anti-Ribonuclease T2 antibody (ab107313) at 1/1000 dilution + NCI-H460 whole cell lysate at 35 µg
Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution