IF staining was performed on HCT15 colon cancer cells using a 1:50 dilution of the primary antibody followed by hybridisation with a 1:100 dilution of the secondary anti-rabbit antibody Alexa Fluor 488 (molecular Probes) with green fluorescence. Picture from Abreview by Karin Birkenkamp-Demtroeder submitted 20 July 2004.
- Showing that non-glycosylated Mrp2 is trapped in the ER in hepatocytes.
Zhang P et al. Role of glycosylation in trafficking of Mrp2 in sandwich-cultured rat hepatocytes. Mol Pharmacol 67:1334-41 (2005). PubMed: 15662046
Mrp2 is a canalicular multispecific anion transport protein in the liver. It is responsible for the biliary excretion of organic anions and is exclusively located on the canalicular membrane. Zhang and his team studied how this localisation was affected by glycolsylation of the protein. They used tunicamycin, a known glycosylation inhibitor to examine its effects on the protein trafficking of Mrp2.
Immunofluorescence studies using Calreticulin antibody (ab4) as an ER marker, showed that when treated with tunicamycin the Mrp2 colocalized with the ER. Since proteins retained in the ER are often subject to proteases this suggested that the non-glycosylated Mrp2 might be degraded by proteasomes. This was supported by western blot analysis.
Antibody data: Rabbit polyclonal
Cross reacts with Cow, Hamster, Human, Mouse and Rat.
Tested in Western blot, Immunoprecipitation, Immunocytochemistry
- Used to identify proteins differentially expressed in different forms of brain tumors.
Odreman F et al. Proteomic studies on low- and high-grade human brain astrocytomas. J Proteome Res 4:698-708 (2005). PubMed: 15952716
Human brain astrocytomas are extremely heterogeneous and difficult to classify on the basis of their microscopic appearance. However, the correct classification is essential for accurate patient prognosis estimates and therapeutic decisions. This paper uses a 2D gel approach to identify 15 proteins that are differentialy expressed in indolent low-grade astrocytomas and high grade malignant gliobastoma multiforme.
PDI was found to be one of the six proteins differentially over-expressed in the low grade astrocytomas. Elevated levels were found mainly in the nuclei of the strocytomas, whereas the high grade tumors were hardly stained. Odreman et al used the Abcam PDI monoclonal in both western blot and immunohistochemical analysis on paraffin-embedded tissues. This paper also used Abcam's alpha Tubulin antibody [DM1A] (ab7291) and Peroxiredoxin 1 antibody (ab15571).
PDI antibody data: mouse monoclonal
Cross reacts with:Human, Mouse, Rat, Dog (see review) and Hamster.
Tested in: Western blot, Immunoprecipitation, Immunohistochemistry (Formalin-fixed paraffin-embedded sections), Immunohistochemistry (Frozen sections), Immunofluorescence, Immunocytochemistry
Other references from 2005:
Balashova N et al. Characterization of a novel type of endogenous activator of soluble guanylyl cyclase. J Biol Chem 280:2186-96 (2005). PubMed: 15509556
Collison LW et al. Aging reduces glycerol-3-phosphate acyltransferase activity in activated rat splenic T-lymphocytes. Biochim Biophys Acta 1687:164-72 (2005). PubMed: 15708364