Immunofluorescence analysis of VAMP1 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 2% BSA for 1 hour at room temperature. The cells were labeled with VAMP1 Rabbit Polyclonal Antibody (PA1765) at 2ug/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) 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||Dog, Human, Mouse, Rat|
|Published species reactivity||Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues M(1) S A P A Q P P A E G T E G A A P G(18) C of rat VAMP-1.|
|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)||0.1-1 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.
|Miscellaneous PubMed (MISC)||See 1 publications below|
PA1-765 detects vesicle associated membrane protein 1 (VAMP-1) from human, canine, mouse and rat tissues.
PA1-765 has been successfully used in Western blot and immunoprecipitation procedures. By Western blot, this antibody detects a 15 kDa protein representing VAMP-1 from rat brain whole protein extract.
The PA1-765 immunogen is a synthetic peptide corresponding to residues M(1) S A P A Q P P A E G T E G A A P G(18) C of rat VAMP-2. This immunizing peptide (Cat. # PEP-099) is available for use in neutralization and control experiments.
The vesicle associated membrane proteins (VAMP) or synaptobrevins are calcium binding proteins specific to eukaryotes. VAMPs, along with synaptosomal associated protein of 25 kDa (SNAP-25) and syntaxin, form the core complex of soluble NSF attachment protein receptor (SNARE) proteins that interact with the soluble proteins N-ethylmaleimide-sensitive factor (NSF) and alpha-SNAP. These membrane associated proteins play a key role in the regulation of vesicle membrane fusion with the plasma membrane. The Clostridium tetani neurotoxin is a metalloprotease with specificity for VAMP. In Alzheimer and quote;s disease, VAMP levels of all isoforms appear to be significantly lowered. It has been shown that the extreme carboxy-terminus of VAMP-1 is involved in subcellular vesicle targeting.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
Tissue expression of the vesicle protein pantophysin.
PA1-765 was used in immunohistochemistry and western blot to study the expression and distribution of the vesicle protein pantophysin
|Windoffer R,Borchert-Stuhlträger M,Haass NK,Thomas S,Hergt M,Bulitta CJ,Leube RE||Cell and tissue research (296:499)||1999|