Western blot analysis was performed on membrane extracts (30 µglysate) of T98G (Lane 1), and RSC96 (Lane 2). The blots were probed with Rabbit Anti-Nicastrin Polyclonal Antibody (Product # PA1-758, 2 µg/ml) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). A 120 kDa band corresponding to immature Nicastrin was observed across the cell lines tested. In addition, a 140 kDa band corresponding to mature, glycosylated Nicastrin was observed in RSC96. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic Peptide: C(688) I N A K A D V L F I A P R E P G A V S Y(708)|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||2 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 9 publications below|
PA1-758 detects nicastrin from human, mouse and rat samples.
PA1-758 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~110 kDa protein representing nicastrin.
PA1-758 immunizing peptide corresponds to amino acid residues 688-708 from human nicastrin. There is a single amino acid substitution between the human and mouse sequence. PA1-758 immunizing peptide (Cat. # PEP-131) is available for use in neutralization and control experiments.
Nicastrin, in addition to presenilin, PEN2, and APH-1 forms the -gamma-secretase protein complex, a membrane-bound aspartyl protease that can cleave certain proteins at peptide bonds buried within the hydrophobic environment of the lipid bilayer. This cleavage is responsible for a key step in signaling from several cell-surface receptors and is thought to be required for the generation of the neurotoxic amyloid peptides that are central to the pathogenesis of Alzheimer and quote;s disease. Like the tumor necrosis factor-alpha-converting enzyme (TACE) and the beta-site cleavage enzyme (BACE) protease families, -gamma-secretase will cleave the amyloid precursor protein (APP), but within the intramembrane region of APP, resulting in either the non-toxic p3 (from the alpha and -gamma cleavage site) or the toxic Abeta amyloid peptide (from the beta and -gamma cleavage site). It is thought that accumulation of the Abeta peptide is the precursor to Alzheimer and quote;s disease. Nicastrin is also thought to be involved in cell proliferation and signaling, especially in regards to activation of Notch receptors as loss of Nicastrin expression results in mouse embryonic lethality.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Presenilin transmembrane domain 8 conserved AXXXAXXXG motifs are required for the activity of the ¿-secretase complex.
PA1-758 was used in western blot to investigate presenilin assembly and catalytic activity
|Marinangeli C,Tasiaux B,Opsomer R,Hage S,Sodero AO,Dewachter I,Octave JN,Smith SO,Constantinescu SN,Kienlen-Campard P||The Journal of biological chemistry (290:7169)||2015|
Presenilins regulate neurotrypsin gene expression and neurotrypsin-dependent agrin cleavage via cyclic AMP response element-binding protein (CREB) modulation.
PA1-758 was used in western blot to study the role of CREB in the mechanism by which neurotrypsin expression and agarin proteolysis is modulated by presenilins
|Almenar-Queralt A,Kim SN,Benner C,Herrera CM,Kang DE,Garcia-Bassets I,Goldstein LS||The Journal of biological chemistry (288:35222)||2013|
Ubiquilin-1 modulates ¿-secretase-mediated ¿-site cleavage in neuronal cells.
PA1-758 was used in western blot to study the ability of a variant of ubiquilin-1 to increase gamma-secretase-mediated cleavage of APP at the epsilon site
|Viswanathan J,Haapasalo A,Kurkinen KM,Natunen T,Mäkinen P,Bertram L,Soininen H,Tanzi RE,Hiltunen M||Biochemistry (52:3899)||2013|
Chronic administration of anti-stroke herbal medicine TongLuoJiuNao reduces amyloidogenic processing of amyloid precursor protein in a mouse model of Alzheimer's disease.
PA1-758 was used in western blot to study the effects of the herbal medicine TongLuoJuiNao on levels of BACE1, gamma secretase components, and beta-amyloid production in a murine model of Alzheimer's disease
|He P,Li P,Hua Q,Liu Y,Staufenbiel M,Li R,Shen Y||PloS one (8:null)||2013|
Calsenilin regulates presenilin 1/¿-secretase-mediated N-cadherin ¿-cleavage and ß-catenin signaling.
PA1-758 was used in western blot to identify the role of Calsenilin in N-cadherin cleavage in neuroblastomas
|Jang C,Choi JK,Na YJ,Jang B,Wasco W,Buxbaum JD,Kim YS,Choi EK||FASEB journal : official publication of the Federation of American Societies for Experimental Biology (25:4174)||2011|
Potent amyloidogenicity and pathogenicity of Aß43.
PA1-758 was used in western blot to investigate the contribution of A beta 43 to Alzheimer disease pathology
|Saito T,Suemoto T,Brouwers N,Sleegers K,Funamoto S,Mihira N,Matsuba Y,Yamada K,Nilsson P,Takano J,Nishimura M,Iwata N,Van Broeckhoven C,Ihara Y,Saido TC||Nature neuroscience (14:1023)||2011|
Ubiquilin 1 modulates amyloid precursor protein trafficking and Abeta secretion.
PA1-758 was used in western blot to investigate the role of ubiquilin 1 during the amyloid precursor protein trafficking and Abeta secretion.
|Hiltunen M,Lu A,Thomas AV,Romano DM,Kim M,Jones PB,Xie Z,Kounnas MZ,Wagner SL,Berezovska O,Hyman BT,Tesco G,Bertram L,Tanzi RE||The Journal of biological chemistry (281:32240)||2006|
The presenilin proteins are components of multiple membrane-bound complexes that have different biological activities.
PA1-758 was used in western blot to study multiple membrane-bound presenilin complexes and their biological activities.
|Gu Y,Sanjo N,Chen F,Hasegawa H,Petit A,Ruan X,Li W,Shier C,Kawarai T,Schmitt-Ulms G,Westaway D,St George-Hyslop P,Fraser PE||The Journal of biological chemistry (279:31329)||2004|
APH-1 interacts with mature and immature forms of presenilins and nicastrin and may play a role in maturation of presenilin.nicastrin complexes.
PA1-758 was used in western blot to investigate the role of APH-1 during the formation of presenilin and nicastrin complexes.
|Gu Y,Chen F,Sanjo N,Kawarai T,Hasegawa H,Duthie M,Li W,Ruan X,Luthra A,Mount HT,Tandon A,Fraser PE,St George-Hyslop P||The Journal of biological chemistry (278:7374)||2003|