1/500 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Application notesIs unsuitable for IP.
FunctionUnknown. Cellular response protein to viral infection.
Tissue specificityWidely expressed. Highly expressed in colon, prostate, trachea and stomach. Expressed at lower level in testis, muscle, lymphoid tissues, white blood cells and spleen. Predominantly expressed by cells of the epithelial lineage. Expressed at low level in normal liver. Expression significantly increases in virus (HBV, HCV) infected liver. Expression does not increase in liver disease due to non-viral causes (alcohol-induced liver disease, autoimmune hepatitis). Increased expression in hepatocytes appears to be a general feature of advanced liver disease. In liver tissue from patients with adult giant-cell hepatitis (GCH), it is strongly expressed in hepatocyte-derived syncitial giant cells. Constitutively expressed by biliary epithelial cells but not by hepatocytes.
Sequence similaritiesBelongs to the GOLM1/CASC4 family.
Post-translational modificationsGlycosylated. Phosphorylation sites are present in the extracelllular medium.
Cellular localizationGolgi apparatus > cis-Golgi network membrane. Early Golgi. Cycles via the cell surface and endosomes upon lumenal pH disruption.
Overlay histogram showing HeLa cells stained with ab109628 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab109628, 1/10000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Western blot - GOLPH2 antibody [EPR3606] (ab109628)
All lanes : Anti-GOLPH2 antibody [EPR3606] (ab109628) at 1/1000 dilution
Lane 1 : LnCaP cell lysate Lane 2 : SH-SY5Y cell lysate
Lysates/proteins at 10 µg per lane.
Secondary HRP labelled Goat anti-Rabbit at 1/2000 dilution
Immunohistochemical analysis of GOLPH2 in paraffin-embedded Human colon tissue using ab109628 at 1/500 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GOLPH2 antibody [EPR3606] (ab109628)This image is courtesy of an anonymous Abreview
ab109628 staining GOLPH2 in Human small intestine by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
Tissue was fixed with paraformaldehyde and blocked with 5% milk for 30 min at 37°C; antigen retrieval was by heat mediation. Samples were incubated with primary antibody (1/1000 in diluent) for 1 hour at 37°C. An undiluted HRP-conjugated goat anti-rabbit polyclonal IgG was used as the secondary antibody.