Immunofluorescence analysis of MUNC18 was performed using 70% confluent log phase U-87 MG 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 MUNC18 Rabbit Polyclonal Antibody (PA1-742) 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 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||Dog, Rat, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic Peptide: V(58) E D I N K R R E P I P S(70)|
|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|
|Immunohistochemistry (Frozen) (IHC (F))||Assay dependent|
|Immunoprecipitation (IP)||Assay dependent|
|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.
PA1-742 detects Munc-18 from human, rat and mouse tissues.
PA1-742 has been successfully used in Western blot, immunohistochemistry, and immunoprecipitation procedures. By Western blot, this antibody detects an ~67 kDa protein representing Munc-18 from rat and mouse whole brain extract.
PA1-742 immunizing peptide corresponds to amino acid residues 58-70 from human Munc-18. This sequence is completely conserved between human, mouse, rat, bovine, and canine Munc-18. PA1-742 immunizing peptide (Cat. # PEP-067) is available for use in neutralization and control experiments.
Munc-18 is a cytosolic brain protein of ~67 kDa which plays a key role in vesicle membrane fusion events with the plasma membrane. Functional interactions have been demonstrated to exist between Munc-18, synaptosomal associated protein of 25 kDa (SNAP-25), synaptobrevin, and syntaxin. Munc-18 is the mammalian homologue of the C. elegans gene unc-18. Mutations in Munc-18 can result in accumulations of acetylcholine and paralytic phenotype suggestive of a neurotransmitter release defect. Munc-18 thus appears to be required by the synaptic vesicle fusion complex. Two Munc-18 isoforms have recently been identified. Munc-18-1 has been shown to be primarily expressed in brain, while Munc-18-2 is expressed in most tissues.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
The Toll-like receptor-3 agonist polyinosinic:polycytidylic acid triggers nigrostriatal dopaminergic degeneration.
PA1-742 was used in western blot to study the role of neuroinflammation in the degeneration of nigrostriatal dopaminergic neurons
|Deleidi M,Hallett PJ,Koprich JB,Chung CY,Isacson O||The Journal of neuroscience : the official journal of the Society for Neuroscience (30:16091)||2010|
A novel ubiquitous form of Munc-18 interacts with multiple syntaxins. Use of the yeast two-hybrid system to study interactions between proteins involved in membrane traffic.
PA1-742 was used in western blot to investigate the interaction between Munc-18 isoforms with syntaxin isoforms
|Hata Y,Südhof TC||The Journal of biological chemistry (270:13022)||1995|
Apical targeting of syntaxin 3 is essential for epithelial cell polarity.
PA1-742 was used in immunoprecipitation to investigate the role of the apical targeting of syntaxin 3 for epithelial cell polarity.
|Sharma N,Low SH,Misra S,Pallavi B,Weimbs T||The Journal of cell biology (173:937)||2006|
Munc18c regulates insulin-stimulated glut4 translocation to the transverse tubules in skeletal muscle.
PA1-742 was used in immunohistochemistry to study the role of Munc18c in GLUT4 translocation in skeletal muscle.
|Khan AH,Thurmond DC,Yang C,Ceresa BP,Sigmund CD,Pessin JE||The Journal of biological chemistry (276:4063)||2001|