Immunofluorescence analysis of SNAP-23 was performed using 90% 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 SNAP23 Rabbit Polyclonal Antibody (PA1-738) 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 membrane 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||Bovine, Mouse, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Sythetic Peptide: C N(193) K N R I D I A N T R A K K L I D S(210)|
|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|
|Immunoprecipitation (IP)||Assay dependent|
|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.
|Western Blot (WB)||See 3 publications below|
PA1-738 detects synaptosomal-associated protein 23 (SNAP-23) from human, mouse and rat samples.
PA1-738 has been successfully used in Western blot and immunoprecipitation procedures. By Western blot, this antibody detects an ~23 kDa protein representing SNAP-23 from rat/mouse brain protein extract and HeLa cell lysates.
PA1-738 immunizing peptide corresponds to amino acid residues 193-210 from mouse SNAP-23 protein. PA1-738 immunizing peptide (Cat. # PEP-147) is available for use in neutralization and control experiments.
Synaptosomal-associated proteins (SNAPs) are cytosolic proteins that play a key role in in the process of membrane fusion in intracellular vesicle trafficking. In eukaryotic cells, the SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) complex is critical to membrane docking and fusion and is believed to impart some degree of specificity between vesicle SNARE (v-SNARE) and target organelle SNARE (t-SNARE). In neurons and neuroendocrine cells, the SNARE complex consists of the integral membrane proteins VAMP (vesicle-associated membrane protein), syntaxin and SNAP-25. In non-neuronal tissue, a SNAP-25 homolog, SNAP-23, functionally replaces SNAP-25 in the SNARE complex. Studies show that VAMP, syntaxin and SNAP-23 are required for SNARE function and that this complex exists as a heterotrimer of the three proteins. In insulin signalling pathways, studies suggest that the translocation of GLUT4 in adipocytes requires that functional SNAP-23 be present in SNARE complexes.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Stimulus-induced S-nitrosylation of Syntaxin 4 impacts insulin granule exocytosis.
PA1-738 was used in western blot to investigate the effect of syntaxin 4 S-nitrosylation on insulin granule exocytosis
|Wiseman DA,Kalwat MA,Thurmond DC||The Journal of biological chemistry (286:16344)||2011|
Developmental changes in the milk fat globule membrane proteome during the transition from colostrum to milk.
PA1-738 was used in western blot to use a shotgun proteomics aproach to study changes in the milk fat globule membrane during the transition from colostrum to milk
|Reinhardt TA,Lippolis JD||Journal of dairy science (91:2307)||2008|
Effects of endurance exercise training on insulin signaling in human skeletal muscle: interactions at the level of phosphatidylinositol 3-kinase, Akt, and AS160.
PA1-738 was used in western blot to investigate the effects of endurance exercise training on insulin-stimulated glucose uptake in human skeletal muscle
|Frøsig C,Rose AJ,Treebak JT,Kiens B,Richter EA,Wojtaszewski JF||Diabetes (56:2093)||2007|