This antibody gave a positive signal in the following tissue lysates: E10 Mouse Embryonic Brain; E12 Mouse Embryonic Brain and Spinal Cord; E16 Mouse Embryonic Brain. ICC/IF: Mouse Embryonic Stem cells.
Storage instructionsShipped 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.
Storage bufferpH: 7.40 Preservative: 0.02% Sodium azide Constituent: PBS Note: Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 120 kDa (predicted molecular weight: 78 kDa).
1/8000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Use a concentration of 5 µg/ml.
FunctionEssential 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). It probably represents a stabilizing cofactor required for the assembly of the gamma-secretase complex.
Tissue specificityWidely expressed.
Sequence similaritiesBelongs to the nicastrin family.
Cellular localizationMembrane. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
ICC/IF image of ab122969 stained Mouse Embryonic Stem 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 ab122969 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nicastrin antibody (ab122969)This image is courtesy of an Abreview submitted by Carl Hobbs
ab122969 staining Nicastrin in Human breast carcinoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/8000 in TBS/BSA/azide) for 16 hours at 21°C. A Biotin-conjugated Goat anti-rabbit IgG polyclonal (1/250) was used as the secondary antibody.
Xu TH et al. Alzheimer's disease-associated mutations increase amyloid precursor protein resistance to ?-secretase cleavage and the Aß42/Aß40 ratio. Cell Discov2:16026 (2016).
Read more (PubMed: 27625790) »