ab18189 specifically detects mouse 12kDa Pen2 protein in lung lysate. A number of higher MW bands are also detected (~50 and ~100kDa bands) which may be because a proportion of Pen2 is found bound to components of the gamma secretase complex (see the Relevance section). All bands detected with ab18189 in mouse lung lysate were blocked with the immunising peptide ab19147. Human Pen2 protein was also detected (weakly) in lung lysates with ab18189 (2ug/ml) in our hands and all detected bands blocked by immunising peptide ab19147.
For endogenous sample testing in (e.g. in lung, spleen or brain tissue), it is suggested that the investigator load a greater amount of protein on the gel and combine immunoprecipitation with Western blot analysis.
This antibody gave a positive result in HeLa cell line in ICC/IF
PEN2 is a functional component of the gamma-secretase complex; coimmunoprecipitation of endogenous PEN2 and PS1 fragments using PS1 antibodies has been reported (Steiner et al, 2002).
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 2 µg/ml. Detects a band of approximately 11.9 kDa (predicted molecular weight: 12 kDa).
Use a concentration of 1 µg/ml.
Essential subunit of the gamma-secretase complex, an endoprotease complex that catalyzes the intramembrane cleavage of integral membrane proteins such as Notch receptors and APP (beta-amyloid precursor protein). Probably represents the last step of maturation of gamma-secretase, facilitating endoproteolysis of presenilin and conferring gamma-secretase activity.
Widely expressed. Expressed in leukocytes, lung, placenta, small intestine, liver, kidney, spleen thymus, skeletal muscle, heart and brain.
Belongs to the PEN-2 family.
Endoplasmic reticulum membrane. Golgi apparatus > Golgi stack membrane. Predominantly located in the endoplasmic reticulum and in the cis-Golgi.
ICC/IF image of ab18189 stained HeLa cells. The cells were 100% methanol fixed (5 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 (ab18189, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit 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.