Immunofluorescence analysis of Fe65 was performed using 70% confluent log phase SH-SY5Y 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 Fe65 Rabbit Polyclonal Antibody (Product # PA1-752) at 2 µg/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, Rat|
|Published species reactivity||Mouse, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic Peptide: P(352) Q E E E K L S Q R N A N P G I K C(369)|
|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 *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Western Blot (WB)||1-2 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.
PA1-752 detects Fe65 from purified mouse samples. This antibody does not detect endogenous levels of Fe65.
PA1-752 has been successfully used in Western blot procedures. By Western blot, this antibody detects an 97 kDa protein representing Fe65 from immunopurified mouse brain lysate.
The PA1-752 immunizing peptide corresponds to amino acid residues 352-369 from mouse Fe65. This peptide (Cat. # PEP-200) is available for use in neutralization and control experiments.
The major transcript of the Fe65 gene is a neuron-specific mRNA that encodes a nuclear protein whose amino-terminal domain strongly activates the transcription of reporter genes. Fe65 has been implicated in a regulatory and cell signaling mechanism because it contains two different motifs involved in protein binding; a WW domain (a variant of Src homology 3 domains) and a phosphotyrosine interaction domain (PID). Interestingly, the PID domain binds to the same motif present in the conserved cytoplasmic domains of the beta PP and beta PP-like proteins.
RNA analyses revealed that Fe65 is predominantly expressed in the brain and in the regions most affected by Alzheimer and quote;s disease-associated neuropathology.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Age-related accumulation of Reelin in amyloid-like deposits.
PA1-752 was used in immunocytochemistry to examine the Reelin accumulation of Reelin in amyloid-like deposits during ageing
|Knuesel I,Nyffeler M,Mormède C,Muhia M,Meyer U,Pietropaolo S,Yee BK,Pryce CR,LaFerla FM,Marighetto A,Feldon J||Neurobiology of aging (30:697)||2009|
RNA interference silencing of the adaptor molecules ShcC and Fe65 differentially affect amyloid precursor protein processing and Abeta generation.
PA1-752 was used in western blot to investigate the effects of RNAi-mediated silencing of the adaptor molecule Fe65 on amyloid precursor protein (APP) processing and A-beta generation
|Xie Z,Dong Y,Maeda U,Xia W,Tanzi RE||The Journal of biological chemistry (282:4318)||2007|