Immunofluorescence analysis of Mint3 was performed using 90% confluent log phase Neuro-2a 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 Mint3 Rabbit Polyclonal Antibody (PA1-072) 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 (A27034) at 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 membranous 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||Rat, Non-human primate, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues M(1) E F L P E P Q H P P G P P T M D L E(19) of rat Mint3.|
|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)||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.
PA1-072 detects munc-18 interacting protein 3 (Mint3) from mouse cells and rat brain samples.
PA1-072 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~86 kDa protein representing Mint3 from AtT20 cell extract and rat brain samples.
The PA1-072 immunogen is a synthetic peptide corresponding to residues M(1) E F L P E P Q H P P G P P T M D L E(19) of rat Mint3. The PA1-072 immunizing peptide (Cat. # PEP-135) is available for use in neutralization and control experiments.
The munc-18 interacting protein (Mint) protein family is a group of evolutionarily conserved adaptor proteins that function in membrane transport and organization. In mammals, there exist three mint isoforms, Mint1, 2, and 3. Although there is little amino acid sequence conservation in the amino-terminal half, the carboxy-terminal half of these proteins is highly conserved. Within this conserved portion there exists a phosphotyrosine-binding (PTB) and a PSD-95/DLG-A/ZO-1 (PDZ) domain, which function as protein interaction modules. Mint1 and 2 appear to be expressed exclusively in the brain and are found to bind to Munc18, an essential component of the synaptic vesicle fusion machinery. Mint3 is ubiquitously expressed in all tissues and is expressed at the lowest levels in the brain and testis. Studies show that mint3 does not interact with munc-18. Mint3 has been found to interact with the Alzheimer and quote;s Disease-related amyloid precursor protein (APP) and does so through its PTB and PDZ domains. It has been suggested that mint3 links APP to other transport machinery components, thereby regulating it transport, endocytosis, and metabolism. Abnormal APP metabolism has been shown to be the cause of an early-onset type of Alzheimer and quote;s disease.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Mint proteins are required for synaptic activity-dependent amyloid precursor protein (APP) trafficking and amyloid ß generation.
PA1-072 was used in immunocytochemistry to study the role of Mint adaptor proteins in the activity-dependent trafficking of APP and PS1 and the generation of beta-amyloid
|Sullivan SE,Dillon GM,Sullivan JM,Ho A||The Journal of biological chemistry (289:15374)||2014|
|Non-human primate||Not Cited||
Intracellular amyloid precursor protein sorting and amyloid-ß secretion are regulated by Src-mediated phosphorylation of Mint2.
PA1-072 was used in western blot to study the role of Src-mediated phosphorylation of Mint2 in the regulation of intracellular APP sorting and amyloid-beta secretion
|Chaufty J,Sullivan SE,Ho A||The Journal of neuroscience : the official journal of the Society for Neuroscience (32:9613)||2012|
Regulation of APP-dependent transcription complexes by Mint/X11s: differential functions of Mint isoforms.
PA1-072 was used in western blot to investigate the differential functions of the Mint isoforms.
|Biederer T,Cao X,Südhof TC,Liu X||The Journal of neuroscience : the official journal of the Society for Neuroscience (22:7340)||2002|
Neuronal expression of mint1 and mint2, novel multimodular proteins, in adult murine brain.
PA1-072 was used in immunohistochemistry to investigate the expression and distribution of mint1 and mint2 in adult murine brain
|Nakajima Y,Okamoto M,Nishimura H,Obata K,Kitano H,Sugita M,Matsuyama T||Brain research. Molecular brain research (92:27)||2001|