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Immunofluorescence analysis of APH1A 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 APH1A Rabbit Polyclonal Antibody (Product # 38-3600) at 2µg/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 (Product # 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 (Product # 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||Rat, Virus, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide derived from the C-terminal region of human Aph-1a|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||Assay Dependent|
* 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.
APH1 is an essential subunit of the gamma-secretase complex, an endoprotease complex that catalyzes the intramembrane cleavage of integral proteins such as Notch receptors and APP (beta-amyloid precursor protein). The gamma-secretase complex is minimally composed of a presenilin homodimer (PSEN1 or PSEN2), nicastrin (NCSTN), APH1 (APH1A or APH1B) and PEN2, although other components may exist. APH1 probably represents a stabilizing cofactor for the presenilin homodimer that promotes the formation of a stable complex.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Mouse||Not Cited||EphB signaling regulates target innervation in the developing and deafferented auditory brainstem.||Nakamura PA,Hsieh CY,Cramer KS||Developmental neurobiology (72:1243)||2012|
|Mouse||Not Cited||Conditional forebrain inactivation of nicastrin causes progressive memory impairment and age-related neurodegeneration.||Tabuchi K,Chen G,Südhof TC,Shen J||The Journal of neuroscience : the official journal of the Society for Neuroscience (29:7290)||2009|
|Transcriptional regulation of PEN-2, a key component of the gamma-secretase complex, by CREB.||Wang R,Zhang YW,Sun P,Liu R,Zhang X,Zhang X,Xia K,Xia J,Xu H,Zhang Z||Molecular and cellular biology (26:1347)||2006|
|Mouse||Not Cited||Nicastrin is critical for stability and trafficking but not association of other presenilin/gamma-secretase components.||Zhang YW,Luo WJ,Wang H,Lin P,Vetrivel KS,Liao F,Li F,Wong PC,Farquhar MG,Thinakaran G,Xu H||The Journal of biological chemistry (280:17020)||2005|
||Nicastrin is critical for stability and trafficking but not association of other presenilin/gamma-secretase components.||Zhang YW,Luo WJ,Wang H,Lin P,Vetrivel KS,Liao F,Li F,Wong PC,Farquhar MG,Thinakaran G,Xu H||The Journal of biological chemistry (280:17020)||2005|
6530402N02Rik; anterior pharynx defective 1 homolog A; APH-1; aph-1 homolog A, gamma secretase subunit; APH-1A; aph-1alpha; APH1A gamma secretase subunit; CGI-78; gamma-secretase subunit APH-1A; presenilin-stabilization factor; RP4-790G17.3
6530402N02Rik; APH-1; APH-1A; APH1A; CGI-78; PSF; UNQ579/PRO1141