Immunofluorescent analysis of APH-1 was performed using 70% confluent log phase A549 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with APH-1 Rabbit Polyclonal Antibody (PA1-2010) at 2 ug/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A27034) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide, KLH conjugated, corresponding to residues C (245) R R Q E D S R V M V Y S A L R I P P E D (265) of human APH-1aL/CT.|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.005% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/mL|
|Immunofluorescence (IF)||2 µg/mL|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||1:1000|
* 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 8 publications below|
PA1-2010 detects APH-1 in human and mouse samples.
PA1-2010 has been successfully used in immunofluorescent, immunoprecipitation, and Western blot procedures. By Western blot, this antibody detects a ~28 kDa protein representing APH-1 in mouse brain and kidney extracts.
The PA1-2010 immunogen is a Synthetic peptide, KLH conjugated, corresponding to residues C (245) R R Q E D S R V M V Y S A L R I P P E D (265) of the C-Terminal domain of the long isoform of APH-1a. This polyclonal antibody has been referred to as, O2C2 in the reference listed above.
Familial Alzheimer and quote;s disease is often characterized by an early (<60 years of age) and rapid deterioration of the central nervous system. The symptoms are caused by the abnormal buildup of senile plaques composed of the 42 residue amyloid-beta peptide. This peptide is the result of the amyloid precursor protein being cleaved by the presenilin-gamma-secretase complex. APH-1 has been found to be a member of the presenilin (PS) complex, and likely contributes to the gamma-secretase activity involved with the onset of Alzheimer's. Recent evidence has shown that APH-1 also plays a role in the maturation of nicastrin/presenilin complexes.
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.
PA1-2010 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|
Familial frontotemporal dementia-associated presenilin-1 c.548G>T mutation causes decreased mRNA expression and reduced presenilin function in knock-in mice.
PA1-2010 was used in western blot to study the effects of a familial frontotemporal dementia-associated presenilin-1 c.548G>T mutation in knock-in mice
|Watanabe H,Xia D,Kanekiyo T,Kelleher RJ,Shen J||The Journal of neuroscience : the official journal of the Society for Neuroscience (32:5085)||2012|
Calsenilin regulates presenilin 1/¿-secretase-mediated N-cadherin ¿-cleavage and ß-catenin signaling.
PA1-2010 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|
Inhibitors of ¿-secretase stabilize the complex and differentially affect processing of amyloid precursor protein and other substrates.
PA1-2010 was used in western blot to study the effect of gamma-secretase inhibitors on amyloid precursor protein
|Barthet G,Shioi J,Shao Z,Ren Y,Georgakopoulos A,Robakis NK||FASEB journal : official publication of the Federation of American Societies for Experimental Biology (25:2937)||2011|
Caspases-2 and -8 are involved in the presenilin1/gamma-secretase-dependent cleavage of amyloid precursor protein after the induction of apoptosis.
PA1-2010 was used in western blot to investigate the involvement of caspase 2 and 8 in the presenilin/gamma-secretase activation and Abeta production in apoptosis
|Chae SS,Yoo CB,Jo C,Yun SM,Jo SA,Koh YH||Journal of neuroscience research (88:1926)||2010|
PI3K inhibition causes the accumulation of ubiquitinated presenilin 1 without affecting the proteasome activity.
PA1-2010 was used in western blot to study the role of PI3K in ubiquitination and proteasome activity
|Aoyagi N,Uemura K,Kuzuya A,Kihara T,Kawamata J,Shimohama S,Kinoshita A,Takahashi R||Biochemical and biophysical research communications (391:1240)||2010|
Presenilins are enriched in endoplasmic reticulum membranes associated with mitochondria.
PA1-2010 was used in western blot to study the expression and localization of presenilins in endoplasmic reticulum membranes
|Area-Gomez E,de Groof AJ,Boldogh I,Bird TD,Gibson GE,Koehler CM,Yu WH,Duff KE,Yaffe MP,Pon LA,Schon EA||The American journal of pathology (175:1810)||2009|
APH-1 interacts with mature and immature forms of presenilins and nicastrin and may play a role in maturation of presenilin.nicastrin complexes.
PA1-2010 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|