Product DescriptionRabbit anti-Nicastrin, Central region Polyclonal Antibody (Unconjugated), suitable for WB, ICC.
Application DetailsWB and IP. Suggested concentration of 3-10 µg/mL is recommended for WB. Human or mouse brain samples commonly prepared with reducing (50 mM DTT), urea (2.3 M), SDS (1.5%) in 62.5 mM Tris-HCL pH 6.8 sample buffer (without boiling) heating to 50 C for 15 min. Unprocessed full length human Nicastrin is 709 amino acids, however this protein contains an N-terminal signal peptide which is considered to undergo removal/cleavage during processing and transit to the cell plasma membrane, in addition the protein undergoes glycosylation to produce a glycoprotein of about 145 kDa apparent MW by SDS PAGE. Biosensis recommends that the optimal working dilution should be determined by the end user.
TargetNicastrin, Central region
SpecificityConfirmed by WB using peptide absorption.
Target Host SpeciesHuman
Immunogen DescriptionA synthetic peptide (C-QGETFDYIGSSRMVYD) corresponding to human Nicastrin [331-346] in the central region conjugated via additional N-terminal Cys to Diphtheria toxoid.
Purity DescriptionProtein G purified IgG
FormatLyophilized from PBS, pH 7.4. Contains no preservative.
Reconstitution InstructionsSpin vial briefly before opening. Reconstitute in 500 µL sterile-filtered 1X PBS, pH 7.2-7.6. Centrifuge to remove any insoluble material.
Storage InstructionsShort term storage at 2-8°C for one week. At -20°C as an undiluted liquid for up to 12 months.
Batch NumberPlease see item label.
Expiration Date12 months after date of receipt (unopened vial).
Scientific BackgroundNicastrin, a type 1 membrane glycoprotein, is an essential component of the gamma secretase complex which is critical for the cleavage of the amyloid precursor protein and other membrane proteins. Nicastrin is widely expressed in different tissue types. This antibody reacts with immature forms of Nicastrin. Detection of higher mol. wt. mature forms is likely to be blocked by glycosylation in this region of the protein.
Western Immunoblotting with nicastrin central region antibody of mouse Nicastrin protein in PS1-deficient cell line and adult mouse brain (right panel) in comparison with C-terminal Nicastrin antibody (Catalog number R-1684-500). Membrane proteins were prepared and loaded as 20 µg protein per lane.
General ReferencesCulvenor, J.G., Ilaya, N.T., Ryan, M.T., Canterford, L., Hoke, D., Williamson, N.A., McLean, C.A., Masters, C.L., and Evin, G. (2004) Characterization of Presenilin complex from mouse and human brain using Blue Native gel electrophoresis reveals high expression in embryonic brain and minimal change in complex mobility with Presenilin mutations. Eur. J. Biochem. 271, 375-385. Ilaya, N.T., Evin, G., Masters, C.L., and Culvenor, J.G. (2004) Nicastrin expression in mouse peripheral tissues is not co-ordinated with Presenilin and is high in muscle. J. Neurochem. 91, 230-23